Papaver somniferum strain with high concentration of thebaine

ABSTRACT

The present invention is directed to an improved poppy straw, concentrate of poppy straw and opium of  Papaver somniferum  for the production of thebaine containing little or no oripavine, codeine or morphine. The present invention also provides plants, stands and seeds of  Papaver somniferum  and methods for the production of thebaine.

CROSS-REFERENCE TO RELATED APPLICATIONS

This is a Divisional application of U.S. patent application Ser. No.12/398,245, filed Mar. 5, 2009, which claims priority to U.S.Provisional Application Ser. Nos. 61/034,583 filed Mar. 7, 2008,61/088,903 filed Aug. 14, 2008 and 61/089,163 filed Aug. 15, 2008, thecontents of each of which are hereby incorporated by reference in theirentirety.

FIELD OF THE INVENTION

The present invention is directed to the improved production ofthebaine. More particularly, the present invention relates to the use ofa mutagenized Papaver somniferum poppy plant to produce thebaine inhigher yield.

BACKGROUND OF THE INVENTION

The 14-hydroxymorphinans, such as, oxycodone, naloxone, naltrexone,naltrexone methobromide, nalbuphine and nalmefene are important opiatederivatives due to their behavior as potent analgesics and/or narcoticantagonists. The most practical synthetic routes to the preparation ofthese pharmaceuticals have utilized the alkaloid, thebaine, as astarting material. Other important opiate derivatives such ashydrocodone and the ring-C bridged compounds buprenorphine and etorphineare also most practically prepared from thebaine.

In accordance with one conventional process, thebaine is oxidized to14-hydroxycodeinone by use of m-chloroperbenzoic acid in an aceticacid/trifluoroacetic acid mixture or by a mixture of hydrogen peroxideand formic acid. 14-hydroxycodeinone is catalytically reduced tooxycodone. See Scheme 1. Oxycodone is a product sold for use as ananalgesic and its production consumes large amounts of thebaine.

Oxycodone can be, in turn, O-demethylated with boron tribromide to yieldoxymorphone. After blocking of the hydroxyl groups with a suitableblocking agent, such as, acetyl groups, the oxymorphone derivative canbe reacted with cyanogen bromide in a von Braun demethylation to yieldan N-cyanodihydronormorphinone derivative that is thereafter hydrolyzedto 14-hydroxydihydronormorphinone (noroxymorphone). Noroxymorphone canbe readily converted to nal-compounds by N-alkylation with appropriatealkyl halide, or acylation with appropriate acyl halide or anhydride,followed by reduction. Another process, converts the oxycodone of theabove process to noroxycodone by the von Braun N-demethylation followedby conversion to a 3-O-methyl-nal-compound using N-alkylation with anappropriate alkyl halide, or by alkylation with an appropriate alkylhalide, or acylation with appropriate acyl halide or anhydride, followedby reduction. The 3-O-methyl-nal-compound is converted to a nal-compoundby O-demethylation.

A synthesis using thebaine to produce the ring-C bridged opiate,buprenorphine, is shown in Scheme 2.

Another synthesis using thebaine to produce the 14-hydroxymorphinan,naltrexone as representative of the nal-compounds, is shown in Scheme 3.

Another synthesis uses thebaine to produce hydrocodone as set forth inU.S. Pat. No. 3,812,132.

Although these syntheses are effective, the availability of thebaine islimited by its high cost. The high cost of thebaine contributes to thehigh cost of the 14-hydroxymorphinans derived from it.

One reason for the limited availability of thebaine, and its high cost,is that total synthesis is difficult. U.S. Pat. Nos. 4,613,668 and4,795, 813 discuss the scarcity of thebaine and teach the totalsynthesis, or alternative synthesis, of the 14-hydroxymorphinans. Yet,the demand for thebaine remains.

A second reason for the limited availability of thebaine, and its highcost, is that the primary source of thebaine is extraction from thepoppy plant, Papaver somniferum. Morphine is the major alkaloid thataccumulates in capsules of Papaver somniferum. Thus, the supply ofthebaine is to a great degree limited to some fraction of the demand formorphine.

Alkaloids are extracted from the poppy capsules of Papaver somniferum bytwo commercial methods. In one method, the immature capsule is cut andthe latex collected from the wound. The air-dried latex is opium which,according to the Merck Index, 11th edition, contains alkaloids in theamounts shown in Table I. In a second method, the mature poppy capsulesand the poppy capsule stems are collected, and threshed to remove theseeds and form a straw. When necessary, the straw is dried to a watercontent below 16%. Solvent or water extraction is employed to remove thealkaloids from the straw. For the varieties of Papaver somniferumnormally grown, the straw, on a dry basis, contains alkaloids in theamounts shown in Table 1.

TABLE 1 opium straw morphine, % 10-16 1-3 codeine, % 0.8-2.5 0.05-0.3 oripavine, %  0-0.1   0-0.05 thebaine, % 0.5-2  0.15-0.65

As can be seen, the yield of thebaine and oripavine is confounded withthat of other alkaloids. A poppy producing predominantly thebaine, e.g.,as 90% or more of the total alkaloids, would enable a simplerextraction/purification process, resulting in higher yields, betterquality and throughput and lower costs.

Where solvent or water or super critical fluid, such as CO₂, extractionis employed to remove the alkaloids from the straw, such method, aspracticed, involves the production of “Concentrate of Poppy Straw”.Concentrate of Poppy Straw (or “CPS”) is described as “The materialarising when poppy straw has entered into a process for theconcentration of its alkaloids, when such material is made available intrade,” (Multilingual dictionary of narcotic drugs and psychotropicsubstances under international control, United Nations, New York, 1983).Not inconsistent with the foregoing description, Concentrate of PoppyStraw is described as “the crude extract of poppy straw in eitherliquid, solid or powder form which contains the phenanthrene alkaloidsof the opium poppy,” 45 U.S. Federal Register 77466, Nov. 24, 1980. Whenin liquid form, the liquid is preferably concentrated before enteringinto commerce. The generally preferred Concentrate of Poppy Straw is thepowder form which results from removing the solvent or water followingextraction of the poppy straw. According to the United Nationspublication “Narcotic Drugs: Estimated World Requirements for 2007;Statistics for 2005 (E/INCB/2006/2)”, Concentrate of Poppy Straw is thedried residue obtained through the extraction of alkaloids from poppystraw. Until the second half of the 1990s, only concentrate of poppystraw containing morphine as the main alkaloid was manufactured. Sincethen, concentrate of poppy straw containing mainly thebaine or oripavinehas started to be manufactured.

More recently, Fist et al., in U.S. Pat. No. 6,067,749 (the “'749Patent”), U.S. Pat. Nos. 6,376,221 and 6,723,894, disclosed an improvedpoppy straw of a stably reproducing Papaver somniferum for theextraction of thebaine and/or oripavine (the “Norman” poppy), thethreshed straw having thebaine and oripavine constituting about 50% byweight or greater of the alkaloid combination consisting of morphine,codeine, thebaine and oripavine. The Norman poppy straw constituted1.68% thebaine, 0.74% oripavine, 0.05% codeine and no morphine as apercent by weight of the dry straw. (See, column 15, table III of the'749 Patent). While this alleviated the limited availability and highcost of thebaine to some extent, the problem of producing oripavineconcurrently with thebaine contributed significantly to the cost ofproducing thebaine.

For many years, the perennial poppy Papaver bracteatum has been proposedas a source of thebaine. Thebaine is the predominant alkaloid in thisspecies, and in selected strains it can be as high as 98% of the totalalkaloids (Palevitch, D and Levy, A 1992 Acta Horticulturae 306, 33-52).Thebaine is present in the roots as well as capsules. Generally twoyears of growth would be required to obtain a good yield of both rootsand capsules. Papaver bracteatum does not alleviate the problem oflimited availability and high cost of thebaine because of its slowgrowth, low capsule yield and the problems with harvesting andprocessing roots.

SUMMARY OF THE INVENTION

The present invention is directed to a poppy straw comprising a poppystraw of a stably reproducing Papaver somniferum having thebaineconstituting about 90% (preferably, about 95%, more preferably, about97%) by weight or greater of an alkaloid combination, and havingoripavine constituting about 10% (preferably, about 5%, more preferably,about 1%, most preferably, about 0.7%) by weight or less of the alkaloidcombination, wherein the alkaloid combination comprises morphine,codeine, thebaine and oripavine; and wherein thebaine constitutes about3.0% or greater of the poppy straw on a dry weight basis. In a preferredembodiment, thebaine constitutes about 3.5% or greater (preferably,about 4.0% or greater, more preferably, about 4.3% or greater) of thepoppy straw on a dry weight basis. In another embodiment, oripavineconstitutes about 0.4% or lower (preferably, about 0.2% or lower) of thepoppy straw on a dry weight basis. In another embodiment of theinvention, oripavine constitutes between 0.01 to 1.0% (preferably,between 0.02% to 0.5%) by weight of the alkaloid combination in thepoppy straw.

There is also provided by the present invention a stably reproducingPapaver somniferum having two genetic traits controlling thethebaine-only characteristic of the plant, one trait being thatdescribed in U.S. Pat. No. 6,067,749, and the second trait regulatingthe step between thebaine and oripavine, resulting in substantially nooripavine. In an embodiment of the present invention is a Papaversomniferum plant having a high thebaine content of over 3% (preferably,over 3.5%, more preferably, over 4.0%, most preferably, over 4.3%) inthe straw wherein the high thebaine content is provided by twoindependent traits, one trait controlling the accumulation of thebaineand oripavine compared with morphine and codeine, and the second traitcontrolling the accumulation of thebaine compared with oripavine. Instill another embodiment of the invention is a Papaver somniferum planthaving a high thebaine content of over 3% (preferably, over 3.5%, morepreferably, over 4.0%, most preferably, over 4.3%) in the straw on a dryweight basis wherein the high thebaine content is provided by twoindependent genetic changes, one genetic change controlling theaccumulation of thebaine and oripavine compared with morphine andcodeine, and the second genetic change controlling the accumulation ofthebaine compared with oripavine.

There is also provided by the present invention a method to improve thethebaine yield of a stably reproducing Papaver somniferum plant, themethod comprising the steps of:

a) exposing at least one poppy seed of Papaver somniferum to amutagenizing agent,

b) growing at least one poppy seed to produce a plant bearing a leaf oran immature poppy capsule, optionally through multiple self-fertilizedgenerations,

c) sampling the leaf or poppy capsule (or any other latex-containingtissue) for the presence of thebaine, oripavine, morphine and codeine,

d) repeating steps a) to c) until a poppy plant of Papaver somniferum isobtained having thebaine constituting about 90% (preferably, about 95%)by weight or greater, and having oripavine constituting about 10%(preferably, about 5%, more preferably, about 1%) by weight or less, ofthe alkaloid combination, wherein the alkaloid combination comprisesmorphine, codeine, thebaine and oripavine; and

e) collect the seed from the plant obtained in “d” and grow anothergeneration of plants to ensure that the subsequent generation stablyreproduces the high thebaine and low oripavine characteristic. In anembodiment of this method, in step d) the poppy plant obtained hasthebaine constituting about 3.0% (preferably, about 3.5%, morepreferably, about 4.0%, most preferably, about 4.3%) or greater of thepoppy straw, on a dry weight basis, and oripavine constituting about0.4% (preferably, about 0.2%) or lower of the poppy straw, on a dryweight basis.

There is also provided by the present invention a stably reproducingPapaver somniferum plant or a stand of a stably reproducing Papaversomniferum plants, in which the production or activity of the enzymeresponsible for the conversion of thebaine to oripavine has beensubstantially inhibited with the result that upon the harvesting oftheir poppy capsules, the poppy plants will yield a poppy straw havingthebaine constituting about 90% (preferably, about 95%, more preferably,about 97%) by weight or greater of the alkaloid combination, and havingoripavine constituting about 10% (preferably, about 5%, more preferably,about 1%) by weight or less of the alkaloid combination, and the poppyplants will yield a poppy straw wherein thebaine constitutes about 3.0%or greater (preferably, about 3.5% or greater, more preferably, about4.0% or greater, most preferably, about 4.3% or greater) of the poppystraw, on a dry weight basis, and oripavine constitutes about 0.4% orlower (preferably, about 0.2% or lower) of the poppy straw, on a dryweight basis.

The present invention also provides a stably reproducing plant ofPapaver somniferum, which upon the harvesting of their poppy capsuleswill yield a poppy straw having thebaine constituting about 90%(preferably, about 95%, more preferably, about 97%) by weight or greaterof an alkaloid combination, and having oripavine constituting about 10%(preferably, about 5%, more preferably, about 1%, most preferably, about0.7%) by weight or less of the alkaloid combination, wherein thealkaloid combination comprises morphine, codeine, thebaine andoripavine; and wherein thebaine constitutes about 3.0% or greater of thepoppy straw on a dry weight basis. The straw of the stably reproducingPapaver somniferum plant of the present invention preferably containsthebaine constituting about 3.5% or greater (preferably, about 4.0% orgreater, more preferably, about 4.3% or greater) of the poppy straw, ona dry weight basis, and oripavine constituting about 0.4% or lower(preferably, about 0.2% or lower) of the poppy straw, on a dry weightbasis. In another embodiment of the invention, oripavine constitutesbetween 0.01 to 1.0% (preferably, between 0.02% to 0.5%) by weight ofthe alkaloid combination in the stably reproducing Papaver somniferumplant.

Also included in the invention is a plant comprising a stablyreproducing plant of Papaver somniferum which upon the harvesting of itspoppy capsules will yield a poppy straw having thebaine constituting atleast 3% (preferably, at least 3.5%) by weight on a dry basis, andoripavine constituting no more than 0.4% by weight on a dry basis ofsaid straw. Preferably, thebaine constitutes at least 4.0% by weight ona dry basis, and oripavine constitutes no more than 0.2% by weight on adry basis of said straw of the plant.

The present invention also provides a plant comprising a stablyreproducing plant of Papaver somniferum which upon the harvesting of itspoppy capsules will yield a poppy straw having thebaine constituting atleast about 3% (preferably, about 3.5%, more preferably, about 4%, mostpreferably, about 4.3%) by weight on a dry basis, and oripavineconstituting about 0.05 to about 0.5% (preferably, about 0.05 to about0.2%) by weight on a dry basis of said straw.

Also included in the invention is a seed of any of the stablyreproducing poppy plants described above; preferably, the seed is aPapaver somniferum seed which is ATCC PTA-9109.

Also included in the invention is a concentrate of poppy straw for theextraction of thebaine comprising a concentrate of poppy straw of any ofthe stably reproducing poppy plants described above.

Another aspect of the invention is an opium for the extraction ofthebaine comprising an opium of any of the stably reproducing poppyplants described above.

Still another aspect of the invention is a poppy straw comprising apoppy straw of any of the stably reproducing poppy plants describedabove.

Another embodiment of the invention is a method for the production ofthebaine which comprises the steps of:

a) harvesting poppy capsules of any of the stably reproducing poppyplants described above to produce a poppy straw; and

b) chemically extracting the thebaine from the poppy straw.

Still another embodiment of the invention is a method for the productionof thebaine which comprises the steps of:

a) collecting and drying the latex of the immature poppy capsules of anyof the stably reproducing poppy plants described above to produce opium;and

b) chemically extracting the thebaine from the opium.

Also included in the invention is a stand of any of the stablyreproducing Papaver somniferum plants described above.

The present invention also provides a poppy straw comprising a poppystraw of a stably reproducing Papaver somniferum having thebaineconstituting at least about 3% (preferably, about 3.5%, more preferably,about 4%, most preferably, about 4.3%) by weight on a dry basis, andoripavine constituting about 0.05 to about 0.5% (preferably, about 0.05to about 0.2%) by weight on a dry basis of said straw.

In the poppy straw, opium, and concentrate of poppy straw of the presentinvention, thebaine preferably constitutes about 96% by weight orgreater of the alkaloid combination and oripavine constitutes about 0.8%by weight or less of the alkaloid combination; more preferably, thebaineconstitutes about 97% by weight or greater of the alkaloid combinationand oripavine constitutes about 0.7% by weight or less of the alkaloidcombination. In the most preferred embodiments of the present invention,there is substantially no oripavine, morphine or codeine in the alkaloidcombination. In another embodiment of the instant invention, thealkaloid combination further comprises salutaridine, reticuline,laudanine, papaverine and noscapine.

The present invention also provides Papaver somniferum plants, andmethods for producing such plants, having poppy straw with substantiallyhigher thebaine content, and substantially lower oripavine content, suchthat thebaine contents in commercially grown and harvested crops areabout 3.0% or greater, preferably, about 3.5% or greater, morepreferably, 4.0% or greater, most preferably, about 4.3% or greater, andoripavine contents are in the order of about 0.4%, preferably, 0.2%, orlower.

Additional embodiments of the present invention provide poppy straw,concentrate of poppy straw and opium, wherein thebaine constitutes about95% by weight or greater of the alkaloid combination and oripavineconstitutes between about 0.01 to about 1.0% by weight (preferably,between about 0.02% to 0.6% by weight, more preferably, between about0.02% to 0.5% by weight) of the alkaloid combination.

Examplifying the invention is a method for producing a poppy plant ofPapaver somniferum having a stably heritable high thebaine content (thatis, the high thebaine content is stably reproducing) and low oripavinecontent versus morphine and codeine content, the method comprising thesteps of:

-   -   a) exposing at least one poppy seed of Papaver somniferum to a        mutagenizing agent,    -   b) growing the at least one poppy seed to produce a plant        bearing a leaf or an immature poppy capsule, optionally through        multiple self-fertilized generations,    -   c) sampling the leaf or poppy capsule for the presence of        thebaine, oripavine, morphine and codeine, and    -   d) repeating steps b) and c) and optionally step (a) until a        poppy plant of Papaver somniferum is obtained which yields a        poppy straw having thebaine constituting about 90% (preferably,        about 95%) by weight or greater of the alkaloid combination        consisting of morphine, codeine, thebaine and oripavine, and        having oripavine constituting about 10% (preferably, about 5%)        by weight or less of the alkaloid combination, and wherein        thebaine constitutes about 3.0% (preferably, about 3.5%, more        preferably, about 4.0%, most preferably, about 4.3%) or greater        of the poppy straw on a dry weight basis.

Preferably, the Papaver somniferum seed exposed to the mutagenizingagent in step (a) of this method is a Papaver somniferum yielding, uponthe harvesting of its poppy capsules, a poppy straw having a thebaineand oripavine content constituting 50% by weight or greater of thealkaloid combination consisting of morphine, codeine, thebaine andoripavine.

In another embodiment of this method, step (b) comprises growing the atleast one poppy seed to produce a plant bearing a leaf or an immaturepoppy capsule, and self-pollinating to produce seed, and taking the seedthereby produced and producing an M2 generation of plants, and step (c)comprises screening the M2 plants and selecting plants which yield apoppy straw having thebaine constituting about 95% by weight or greaterof the alkaloid combination consisting of morphine, codeine, thebaineand oripavine, and having oripavine constituting about 5% by weight orless of the alkaloid combination, each on a dry weight basis, andwherein thebaine constitutes about 3.5% (preferably, about 4.0%, morepreferably, about 4.3%) or greater of the poppy straw on a dry weightbasis,

In another aspect of this method, the Papaver somniferum seed exposed tothe mutagenizing agent in step (a) of this method is seed selected fromATCC PTA-9110 or ATCC PTA-9109.

The present invention is also directed to progeny of Papaver somniferumATCC-9109, said progeny yielding a poppy straw having thebaineconstituting about 90% (preferably, about 95%) by weight or greater ofan alkaloid combination, and having oripavine constituting about 10%(preferably, about 5%) by weight or less of the alkaloid combination,and wherein thebaine constitutes about 3.0% (preferably, about 3.5%,more preferably, about 4.0%, most preferably, about 4.3%) or greater ofthe poppy straw on a dry weight basis, wherein the alkaloid combinationcomprises morphine, codeine, thebaine and oripavine.

Another example of the present invention is a mutant or variant ofPapaver somniferum ATCC PTA-9109 or ATCC PTA-9110, said mutant orvariant yielding a poppy straw having thebaine constituting about 90%(preferably, about 95%) by weight or greater of an alkaloid combination,and having oripavine constituting about 10% (preferably, about 5%) byweight or less of the alkaloid combination, each on a dry weight basis,and wherein thebaine constitutes about 3.0% (preferably, about 3.5%,more preferably, about 4.0%, most preferably, about 4.3%) or greater ofthe poppy straw on a dry weight basis, wherein the alkaloid combinationcomprises morphine, codeine, thebaine and oripavine.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 provides a chromatogram showing separation of the alkaloids usingthe isocratic UPLC method.

FIG. 2 provides a chromatogram of the poppy straw of the M3 generationof FN1-1242-3. ATA indicates thebaine, on indicates oripavine peak, AMArepresents morphine, and ACA represents codeine.

DETAILED DESCRIPTION OF THE INVENTION

Utilizing the mutagenized plants of Papaver somniferum as describedherein, persons skilled in the art easily know how to grow them,reproduce them, collect the latex or the dried straw and purify thethebaine. As one enablement of the present invention, seeds to themutagenized plants of Papaver somniferum (FN1-1242-3), as describedherein, have been deposited under the Budapest Treaty with the AmericanType Culture Collection, 10801 University Boulevard, Manassas, Va.20110-2209, on Mar. 20, 2008, under ATCC® Patent Deposit DesignationPTA-9109. All restrictions on access to these deposits will beirrevocably removed at the time a patent issues in the United States onthe basis of this application. The availability of these seeds is not tobe construed as a license to practice this invention in contravention ofrights granted under the authority of any government in accordance withits patent or breeder's rights laws. Regardless of the enablementprovided by this deposit, persons skilled in the art of mutagenizingseed, can obtain the seed herein by employing the mutagenesis process asdescribed below.

The production of mutagenized seed is well known in the art. Methods ofseed mutagenesis as well as mutagens suitable for use in these methods,such as ethyl methanesulfonate (EMS), are described in the Manual onMutation Breeding, 2nd ed., I.A.E.A., Vienna 1977 or in Plant Breeding,Principles and Prospects, Chapman and Hall, London 1993. For X-raymutagenized seeds, hydrated seeds might be treated with 20,000 rads, (30cm from the source for 45 minutes using a filter). X-ray mutagenesis isdescribed and compared to EMS mutagenesis by Filippetti, A. et al.,“Improvement of Seed Yield in Vicia Faba L. By Using ExperimentalMutagenesis II Comparison of Gamma-Radiation andEthyl-Methane-Sulphonate (EMS) in Production of Morphological Mutants”,Euphytica 35 (1986) 49-59. DEB, diepoxybutane, mutagenized seeds mightbe obtained by soaking the seeds in water overnight, then soaking in 22mM DEB for 4 hours, followed by extensive washing. Further mutagensinclude ethyl-2-chloroethyl sulphide, 2-chloroethyl-dimethylamine,ethylene oxide, ethyleneimine, dimethyl sulphonate, diethyl sulphonate,propane sulphone, beta-propiolactone, diazomethane,N-methyl-N-nitrosourethane, acridine orange and sodium azide.

Mutagenesis utilizing EMS is well described in the literature. TheManual on Mutation Breeding, supra, reports a preferred EMS mutagenesisprocess for barley seeds as practiced by K. Mikaelson. In this preferredprocess, the seeds are prepared, pre-soaked, treated with the mutagenand post-washed.

U.S. Pat. No. 6,067,749, incorporated by reference herein in itsentirety, describes the use of EMS for the preparation of a Papaversomniferum strain with a high concentration of thebaine and oripavine.

Irradiation methods such as fast neutron mutagenesis may also be used toproduce mutagenized seed. (See, Li, X. et al., A fast neutron deletionmutagenesis-based reverse genetics system for plants, The Plant Journal27 (3), 235-242 (2001)). Applicants employed and prefer fast neutronmutagenesis (“FNM”) as the mutagen herein.

Fast neutron mutagenesis is described by Kodym and Afza (2003), Physicaland Chemical Mutagenesis, pp 189-203, in Methods in Molecular Biology,vol 236: Plant Functional Genomics: Methods and Protocols (Ed. E.Grotewold), Humana Press Inc, Totowa, N.J.

Gamma (γ) Rays are electromagnetic waves of very short wavelengths andare obtained by disintegration of radioisotopes Co or Cs. γ sources canbe installed in a γ cell, a γ room, or γ field. These are shielded bylead or concrete. Most γ sources as suitable for seed irradiation, aslong as the size of irradiation space is sufficient and the dose rateallows practical irradiation times.

Fast neutrons are uncharged particles of high kinetic energy and aregenerated in nuclear reactors or in accelerators. The scientist shouldassess the feasibility for seed irradiation with the operators, sincenot all facilities are suitably equipped and can produce fast neutronsat a low degree of contamination with other radiation.

The two radiation types differ in their physical properties and, hence,in their mutagenic activity. γ Rays have a lower relative biologicaleffectiveness (RBE) than fast neutrons, which implies that in order toobtain the same biological effect, a higher dose of γ radiation must begiven. RBE is mainly a function of the linear energy transfer (LET),which is the transfer of energy along the ionizing track. γ Rays producea few ionizations per micron of path (low LET) and belong to thecategory of sparsely ionizing radiation. Fast neutrons (high LET,densely ionizing radiation) impart some of their high kinetic energy viacollisions, largely with protons within the material.

When radiation passes through tissue, physical events such asionizations (ejection of electrons from molecules) and excitations(process of raising electrons to a higher energy state) occur and leadto effects in the DNA, membranes, lipids, enzymes, etc. Secondly,chemical events are induced that start with the formation of activatedmolecules, so-called free radicals (OH. and H.) that arise from OH− andH+. If oxygen is present, it reacts readily with radiation-induced freeradicals to form peroxyradicals. In the case of low LET radiation, theformation of peroxyradicals is favoured. In high LET radiation, theformation of hydrogen peroxide (H₂O₂) by recombination of free radicalsis favoured. All radicals and hydrogen peroxide can react withbiological molecules. Primary damage caused by radiation occurs randomlyand is both physiological and genetic. Physiological recovery and repairof DNA are possible to some extent, as non-damaged molecules may takeover metabolic processes and DNA repair mechanisms are activated.

Before starting any mutation induction studies, it is most crucial toselect suitable doses. For mutation induction, it is advisable to usetwo to three doses along with a control. The applicable doses willdepend on the breeding or research objective, the radiation type and theparticular plant material. It is known that plant genera and speciesand, to a lesser extent, cultivars differ in their radiosensitivity.Radiosensitivity (radiation sensitivity) is a relative measure thatgives an indication of the quantity of recognizable effects of theradiation exposure on the irradiated object. The radiosensitivity isinfluenced by biological factors (such as genetic differences, nuclearand interphase chromosome vol) and by environmental modifying factors(oxygen, water content, post-irradiation storage, and temperature).

Modifying factors greatly affect mutagenic efficiency andreproducibility of results. Oxygen is the major modifying factor, whilemoisture content, temperature, and storage appear to be secondary,interacting with the oxygen effect. Oxygen shows synergistic action withsparsely ionizing radiation, but oxygen effects during irradiation andpost-irradiation storage can easily be prevented by adjustment of seedwater content to 12-14% in cereals and most other seeds. In oilseedssuch as poppies, the seed water content should be lower, around 7-8%.The critical region is the embryo, but it can be assumed that the watercontent of the seed and the embryo of most species will be similar.Environmental factors are less important with densely ionizingradiation; thus, for fast neutron radiation, no seed moisture adjustmentis necessary.

Unless data on the radiosensitivity of a given plant are alreadypublished or known from experience, the mutation induction programshould be preceded by a radiosensitivity test. This is done byirradiating the seeds with a range of doses and by growing out theplants under greenhouse conditions. Radiosensitivity is assessed basedon criteria such as reduced seedling height, fertility, and survival inthe M1 generation. A seedling height reduction of 30-40% is generallyassumed to give a high mutation yield. The usefulness of radiation canbe judged by mutagenic efficiency, which is the production of desirablechanges free from association with undesirable changes. A high dose willincrease mutation frequency (the frequency at which a specific kind ofmutation or mutant is found in a population of cells or individuals),but will be accompanied by negative features, such as sterility. Whenselecting the doses, it will be necessary to find a treatment regimeproviding high mutagenic efficiency.

For fast neutron radiation, dosimetric measurements have to be doneduring each radiation treatment, e.g., by performing the sulphurthreshold detector method, since the neutron flux in the seedirradiation unit is not constant.

The Gray (symbol Gy), the SI (Systéme Internationale) unit used toquantify the absorbed dose of radiation (1 Gy=1 J/kg) replaced the oldunit rad; 1 Gy=100 rads or 1 krad=10 Gy. The absorbed dose rate (Gy/s orGy/min) indicates how much energy the irradiated material absorbs duringa given unit of time. The length of exposure and the dose ratedetermines the radiation dose. Exposure during short times (s to a fewh) at a high dose rate is referred to as acute and is most applied inirradiation programs.

We used the Atomic Energy Research Institute, Konkoly Thebe ut 29/33,X.epulet, H-1121 Budapest, Hungary to irradiate our seeds.

Fast neutrons have been shown to be a very effective mutagen. Kornneefet al. (1982) found that about 2500 lines treated with fast neutron at adoes of 60 Gy are required to inactivate a gene once on average(Koornneef, M., Dellaert, L. W. M. and van der Veen, J. H. (1982) EMS-and radiation-induced mutation frequencies at individual loci inArabidopsis thaliana (L.) Heynh. Mutat. Res. 93, 109-123). If the plantgenome contains about 25000 genes, it is estimated that about 10 genesare randomly deleted in each line.

FNM offers a number of advantages over using chemical treatment such asEMS. Notably, the treatment is applied to the dried seed, which can besown at a later date, while with EMS the seed needs either to be sownimmediately after treatment, or carefully re-dried for sowing later.

After the seeds have been exposed to the mutagen, the seeds are grown tomaturity in controlled conditions and self-pollinated. The seeds fromthe mature plant are taken and at least one seed is planted to grow anM2 generation. The M2 generation is screened for alkaloid production. Ofcourse, it is possible to screen the M1 generation, but there areseveral advantages to screening the M2 generation. Firstly, screeningthe M2 generation insures that the trait resulting from mutagenesis canbe inherited. Secondly, by growing the M2 generation, the basichardiness of the plant is proven before screening. Thirdly, traitsresulting from mutagenesis are generally inherited as recessive genes.Typically the mutated gene will be in the heterozygous state in the M1generation, and thus the mutation will be masked by the dominant(non-mutated) form of the gene. In the M2 generation, however, in aproportion of the plants the gene will be in the homozygous state, andthe affect of the mutation apparent. The M2 plants can be grown toproduce an immature capsule, but it is possible to save time and laborif the plants are screened at an earlier stage of growth. It isrecommended that the plants be screened at a point beginning at the 6leaf stage, up to the 10 leaf stage. Screening at this early stageallows many plants to be managed in a small space. The screening processitself is the most labor intensive. Thus, to improve return on labor,only plants that appear healthy should be screened.

In the screening process, the objective is to measure each plant formorphine, codeine, thebaine and oripavine content. Additional alkaloidswhich can also be measured during the screening process includesalutaridine, reticuline, laudanine, papaverine and noscapine. This canbe accomplished by extracting, for example, a dry leaf into a liquidbuffer or by dissolving a latex sample into a buffer. The buffersolutions are placed onto 96 well trays and fed mechanically through anyof the high-throughput HPLCs available on the market. In a preferredembodiment, an isocratic Ultra high performance Liquid Chromatography(UPLC) method, as described herein, is utilized which provides veryrapid latex screening.

Plants with interesting alkaloid contents are grown further and examinedin more detail. According to the procedure herein, a second sample istaken from about 3% of plants to clarify or confirm the results of theinitial screen. A more precise gradient UPLC method as described hereinis used to obtain more accurate peak identification and quantification.Plants confirmed to have an unusual alkaloid profile are transplanted to200 mm (approx 8 inch) pots for growing to maturity. Twenty one plantshaving high thebaine and substantially no oripavine, morphine or codeinewere found after screening approximately 34,358 plants.

As used herein, the term “poppy straw” or “straw” shall mean the strawmaterial which results when the mature poppy capsules and the poppycapsule stems of a Papaver somniferum plant are collected, and threshedto remove the seeds to form a straw.

The term “opium”, as used herein, shall refer to the air-dried, milkyexudation (i.e., the latex) from incised, unripe poppy capsules of aPapaver somniferum plant.

As used herein, the term “concentrate of poppy straw” or “CPS” shallmean the material arising when poppy straw has entered into a processfor the concentration of its alkaloids in either liquid, solid or powderform which contains the phenanthrene alkaloids of the opium poppy.

The phrase “stand of Papaver somniferum” or “stand of stably reproducingPapaver somniferum”, as used herein, refers to a group of two or morePapaver somniferum plants or stably reproducing Papaver somniferumplants located together.

As used herein, the term “alkaloid combination” shall refer to acombination of alkaloids wherein the alkaloid comprises morphine,codeine, thebaine and oripavine. In another embodiment of the presentinvention, the alkaloid combination further comprises salutaridine,reticuline, laudanine, papaverine and noscapine in addition to morphine,codeine, thebaine and oripavine.

A “stably reproducing” Papaver somniferum poppy plant as describedherein refers to a poppy plant that is stably reproducing as required toplant and harvest seed poppy crop over multiple generations where eachgeneration would be suitable, without seed selection, for commercialplanting of a field crop or stand of plants exhibiting the desiredalkaloid characteristics. A stably reproducing poppy plant contains thedesired alkaloid characteristics as described herein, and when selfpollinated, or cross pollinated by a plant with the same genescontrolling alkaloid content, produces a subsequent generation of plantswhich substantially all have seed that when grown produces plants withthe same desired alkaloid characteristics as the parent plant. Moreover,in the absence of pollination with pollen from other chemotypes (e.g.,conventional morphine accumulating plants), the line will continue toproduce similar plants over multiple generations, without the need forselection to maintain the desired alkaloid characteristic. An example ofa desired alkaloid characteristic which can be passed on to futuregenerations by a stably reproducing Papaver somniferum poppy plantincludes the improved thebaine characteristics (e.g., wherein thebaineconstitutes about 90% (preferably, 95%, more preferably, 96% and mostpreferably, 97%) by weight or greater, and oripavine constitutes about10% (preferably, 1%, more preferably, 0.8% and most preferably, 0.7%) byweight or less of the alkaloid combination).

As used herein, the “M1 population” is the seeds and resulting plantsexposed to a mutagenic agent, while “M2 population” is the progeny ofself-pollinated M1 plants, “M3 population” is the progeny ofself-pollinated M2 plants, “M4 population” is the the progeny ofself-pollinated M3 plants, and generally “Mn population” is the progenyof self-pollinated Mn-1 plants.

As stated above, there is obtained by the present invention, a poppystraw, concentrate of poppy straw or opium having thebaine constitutingabout 90% by weight or greater and oripavine constituting about 10% byweight or less of an alkaloid combination comprising morphine, codeine,thebaine and oripavine. Preferably, thebaine constitutes about 95% (morepreferably, about 96% and most preferably, about 97%) by weight orgreater, and oripavine constitutes about 1% (more preferably, about 0.8%and most preferably, about 0.7%) by weight or less of the alkaloidcombination comprising morphine, codeine, thebaine and oripavine. In apreferred embodiment, the alkaloid combination further comprisessalutaridine, reticuline, laudanine, papaverine and noscapine. Morepreferably, there is substantially no oripavine, morphine or codeine inthe alkaloid combination, and most preferably, there is substantially nooripavine, morphine, codeine, salutaridine, reticuline, laudanine,papaverine or noscapine in the alkaloid combination.

As used herein, the term “substantially no” when referring to oripavinecontent means that oripavine constitutes less than 0.6% by weight,preferably, less than 0.5% by weight, more preferably, less than 0.4% byweight, and most preferably, between 0% and 0.2% by weight of thealkaloid combination of the poppy straw, concentrate of poppy straw oropium.

The term “substantially no”, when referring to morphine, codeine,salutaridine, reticuline, laudanine, papaverine or noscapine, as usedherein, means that each of the specified alkaloids constitutes less than1% by weight, preferably, less than 0.5% by weight, more preferably,less than 0.3% by weight, and most preferably, between 0% and 0.2% byweight of the alkaloid combination of the poppy straw, concentrate ofpoppy straw or opium.

The term “trait”, as used herein, mean a distinct heritable phenotypiccharacteristic. The desired traits, i.e., high thebaine content versusoripavine, morphine or codeine content, once established are highlyheritable. To maintain the desired traits, care should be taken toprevent cross-pollination with normal plants unless suchcross-pollination is part of a controlled breeding program.

The desired traits can be transferred into poppy lines having othercharacteristics (e.g. different height, early or late maturity or havingdisease resistance) by cross pollinating the high thebaine plant withthe second parent plant, collecting F1 seed, growing a F1 plant which isallowed to self-pollinate and collect the F2 seed. The F2 seed wouldthen be grown, and individual plants that have the high thebainecharacteristic could be selected according to the methods herein, alongwith the other desired characteristics such as disease resistance. Askilled operator will be able to apply variations to this method asknown in plant breeding.

Conducting test crosses with plants of known genotype can provideinformation regarding the genetic changes introduced through mutation.The characteristics of the F1 generation produced by crossing to anormal (wild type) parent will indicate whether a trait inherits as arecessive or dominant gene. Self pollinating the F1 plants anddetermining the phenotypes of the subsequent F2 population of plantswill provide information regarding the numbers of genes responsible forparticular characteristics.

The theory whereby mutagenesis has been found to be capable of raisingthe thebaine content of Papaver somniferum relative to the oripavine,morphine and codeine content is not capable of a certain or definiteexplanation at this time. The mutagenesis might have modified thebiosynthesis pathway in any number of ways to minimize the production oforipavine. Despite the fact that definite answers are not now available,there are good reasons to believe that the correct answer is known.

Papaver somniferum is postulated to have two biosynthetic pathways fromthebaine to morphine as shown in Scheme 4. Pathway A via neopinone,codeinone and codeine was proposed by Parker, H. I., J. Am. Chem. Soc.,94, 1276-1282 (1972). Pathway B via oripavine and morphinone wasproposed by Brochmann-Hanssen, E., Planta Med., 50, 343-345 (1984). Theenzyme codeinone reductase (NADPH) is believed to be active in bothpathways, reducing codeinone to codeine and morphinone to morphine.Further, the TOP1 mutation (Millgate et al., Nature, Vol. 431, 413-414,2004) affects both pathways, preventing thebaine being converted toneopinone in Pathway A, and preventing oripavine being converted tomorphinone in Pathway B. The TOP1 mutation appears to blockdemethylation of the enol ether which converts thebaine to neopinone, aswell as the demethylation of the same enol ether in oripavine.

By the methods herein, plants of Papaver somniferum were obtained havingsubstantially no oripavine, morphine or codeine. Both Pathway A andPathway B were inoperative to produce morphine in the parent line usingthe TOP1 mutation. The most probable step that has been affected bymutation is the phenolic O-demethylation step between thebaine andoripavine. Thus, it is believed, for the Papaver somniferum plantsdescribed herein, that the production or activity of the phenolicO-demethylase enzyme that converts thebaine to oripavine has beensubstantially inhibited. Stably reproducing Papaver somniferum inaccordance with the present invention may also be obtained byrecombinant DNA techniques. In particular, after isolation andsequencing of the gene coding for thebaine demethylase, the gene may bemodified or deleted to inhibit or prevent the production of thebainedemethylase. Techniques for modifying gene activity such as RNAi,antisense and other techniques are well known to those skilled in theart. Once the gene coding is established, a TILLING technique may beused to more efficiently recover mutants from populations (Henikoff, S.,Till, B. J. and Comai, L. (2004) TILLING. Traditional mutagenesis meetsfunctional genomics. Plant Physiology 135, 630-636).

Knowing that there are genetic means of reducing the conversion ofthebaine to oripavine in poppies, and that now that we have shown thatthese poppies are achievable and viable, even conventional breedingapproaches may ultimately be used to develop such plants.

Recovering thebaine from either the dried straw or from the opium ofPapaver somniferum is a process well established in the art. Until now,thebaine has been extracted from this plant species either as a part ofthe process of extracting morphine and codeine, or more recently as partof the process of extracting thebaine and oripavine.

In one process, the straw is treated with a small amount of lime andwater to soften the capsules and to form a free base of the alkaloids.Countercurrent extraction of the softened straw with methanol, ethanolor other suitable solvent forms a solvent/water extract or “miscella”containing the alkaloids, with morphine at a concentration of about 1g/L where the straw is from standard Papaver somniferum. The volume ofthe miscella is reduced about 30× under vacuum to produce an aqueousconcentrate. Thebaine is extracted from aqueous concentrate using aliquid/liquid extraction with toluene, adjusting pH for the bestseparation of thebaine. The thebaine is recovered from the toluene. Ofcourse, recovering thebaine from the improved Papaver somniferumprovided herein will be facilitated by the fact that the concentrationof the thebaine in the miscella will be much higher than that of otheralkaloids and thus can be more easily collected by precipitation. Also,in the substantial absence of oripavine, morphine and codeine, thethebaine might be directly extracted from the straw using toluene,xylene or other organic solvent in which thebaine has solubility.

The following Examples are set forth to aid in the understanding of theinvention, and are not intended and should not be construed to limit inany way the invention set forth in the claims which follow thereafter.

Example 1

A selection of Papaver somniferum poppy, WF03-0802 is used as thestarting material. This line contains the TOP1 mutation and thereforehas the characteristics of containing thebaine and oripavine in itspoppy straw and opium, and is substantially free of morphine andcodeine. Seeds of WF03-0802 have been deposited under the BudapestTreaty with the American Type Culture Collection, 10801 UniversityBoulevard, Manassas, Va. 20110-2209, on Mar. 20, 2008, under ATCC PatentDeposit Designation PTA-9110, and will be made available upon thematuration of this application into a patent. All restrictions to thedeposited seeds will be irrevocably withdrawn for the enforceable lifeof the patent. The availability of these seeds is not to be construed asa license to practice this invention in contravention of rights grantedunder the authority of any government in accordance with its patent orbreeder's rights laws.

Six seed samples each of 10 g were prepared. One sample was retained asa control. After obtaining the necessary inspections and permits, the 5samples were sent to the Atomic Energy Research Institute, Budapest,Hungary for irradiation. At the Institute, the samples were removed fromtheir vials, packed into plastic bags and cadmium holders and irradiatedwith fast neutrons. The dose rates and exposure times were as follows:

-   Treatment 1 10 Gy 13 minutes 17 seconds-   Treatment 2 20 Gy 26 minutes 30 seconds-   Treatment 3 25 Gy 33 minutes 16 seconds-   Treatment 4 35 Gy 46 minutes 27 seconds-   Treatment 5 50 Gy 66 minutes 13 seconds

The reported parameters of the irradiation were as follows:

-   -   Irradiation geometry at BIF of BRR at AERI: 2Y/Cd, rotated    -   Monitored by U-235, Th-232 fission chambers and GM counter.    -   The dose homogeneity within one package is better than 1%    -   The overall dose uncertainty is less than 5%.    -   Total surface gamma activity of samples after irradiation: ˜695        BGND    -   Total surface gamma activity of samples on 20 Oct. 2005: <2 BGND    -   (1 BGND (background) is ˜90 nGy/h)

The seeds were returned to applicants by the Institute, reweighed, andfound to have lost an average mass of 1.1%. This is compatible withbeing transferred into and out of plastic bags, as well as losing somemoisture during irradiation.

Samples of the seeds were placed on damp filter paper in a Petri dish.After 3 and 7 days, the seeds were examined. Treatments 1 to 4germinated well, while treatment 5 had very short radicles and only asmall percentage of plants with shoots.

A pot trial was grown to evaluate the effect of the FNM treatment onplant growth. Two pots were sown with 10 seeds from each of treatments1, 2, 3, 5 and control. These pots were observed for emergence and plantgrowth. Irradiation reduced plant emergence and survival from a mean of9 plants per pot in the control treatment to 8.5 plants in Treatment 1,7.5 plants in treatment 2, 7 plants in treatment 3, and 4 plants intreatment 5. Development was also delayed with increasing dose, andplant height was reduced by about 10% in treatment 1, 20% in treatment2, and 40% in treatment 3.

The seeds from the treatments 2 and 3 were sown into 200 mm pots filledwith potting mix from Forestry Commission Nursery at Perth, Tasmania. 10seeds were sown per pot, and were covered with vermiculite. The plantswere grown through to maturity in a greenhouse. All flowers were selfpollinated by transferring pollen from the anthers onto the stigmaticdisc. The mature capsules were harvested into large paper bags, labeledwith the treatment number, keeping the different treatments separate.Where there were 2 or more capsules on one plant, these were picked intoa paper bag so they stayed together. Distinctive plants were harvestedinto separate bags and notes made on their appearance. The harvestedcapsules were stored for a week or so to ensure that the seed was airdry.

The seed was separated from the capsules in the laboratory, and weighedinto paper envelopes labeled with FN1-X where FN1 refers to Fast Neutronexperiment 1, and X is the sequence number of the seed sample. The seedfrom multiple capsules from the same plant was combined into the onesample.

The seed from a total of 8,495 plants was harvested. 7,280 of these werefrom radiation treatment 2. The median weight of seeds harvested per M1plant was 0.41 g.

Growth and Screening of M2 Generation

Plant Growth

M2 plants were grown in a greenhouse in trays each with 288 cells. 12cells were sown with seed from each M1 plant. Two seeds were sown ineach cell, and thinned to one plant per cell after 1-2 weeks. The plantswere sown in batches of 4-17 trays each week to spread the workload over17 weeks.

Leaf Sampling

When the plants were approximately 6 weeks old, they were analysed foralkaloid profile using the latex from the youngest fully expanded leaf(YFEL). 240 μL of latex extraction solution (23 g NH₄H₂PO₄ dissolved in800 mL deionized (DI) water, made up to 1 L with ethanol) was added tothe wells of 96 well filter plates (Pall AcroPrep™ 96 Filter Plate 0.2μm GHP membrane, natural housing, 350 μL PN S5045, (Pall Corporation,East Hills, N.Y.)). The tip of the YFEL was removed from each plant andplaced in a well of the filter plate using fine forceps. Three filterplates were required to sample the plants in one tray. The plates wereallowed to stand for about 30 minutes after sampling to allow the latexto bleed out of the leaves into the extraction solution. The solutionwas then filtered into a 96 well collection plate, which was sealed withan ABgene® Adhesive PCR foil seals (Abgene, part of ThermoFisherScientific, Rockford, Ill.) to eliminate evaporation.

UPLC Method

The UPLC method used for the first screening stage is described inExample 2. Peak areas were exported to a Excel file (MicrosoftCorporation, Seattle, Wash.) for data analysis. No correction wasapplied for differing UV absorption between the alkaloid peaks. Therelative absorption of oripavine and thebaine, the main peaks ofinterest, were in any case very similar at the wavelength used.

Data Analysis

Relative peak areas were calculated for all identified alkaloids. TheExcel data files were then sorted to identify plants having highthebaine content and low oripavine content relative to all identifiedalkaloids extracted. The chromatograms of plants identified as being ofinterest were reviewed to ensure that the peaks of interest werecorrectly integrated.

Confirmation

The plants identified in the first screening were then resampled toprovide confirmation of the alkaloid profile, and to ensure that thecorrect plant was located prior to transplanting. The selected plantswere marked with a plastic coated wire when retested so that they couldbe identified reliably for transplanting. A gradient UPLC system with a2.5 minute run time (described in Example 2) was used in theconfirmation testing in order to obtain more accurate peakidentification and integration.

Transplanting

Plants confirmed as being of interest were transplanted into 200 mmpots, and labelled with a code, indicating the M1 seed line from whichthey were derived. For instance, if two selections were made from the M1seed sample labelled FN1-1234, these selections were labelledFN1-1234-1, and FN1-1234-2. Up to 5 plants were transplanted into eachpot.

Table 2, below, shows the number of plants analysed, the number ofselections made, and the number of selections confirmed. Over theproject, 34,358 M2 plants (from 4,176 M1 lines) were tested, and 1,049were selected for further testing. 549 of these were confirmed andtransplanted into pots. Of the 549 transplanted, 366 were selected onthe basis of high thebaine and low oripavine content.

TABLE 2 Irradiation Number Confirmations Treatment Tray plantsSelections % of % of Batch No. numbers analysed No. % selns No. plantsselections 1 2 1-4 927 51 5.5 23 2.5 45 2 2 5-8 856 15 1.8 12 1.4 80 3 2 9-12 914 29 3.2 23 2.5 79 4 2 13-16 976 37 3.8 27 2.8 73 5 2 17-20 92440 4.3 14 1.5 35 6 2 21-28 1900 47 2.5 21 1.1 45 7 2 29-35 1670 27 1.616 1.0 59 8 2 36-43 1746 85 4.9 30 1.7 35 9 2 44-52 2134 39 1.8 39 1.8100 10 2 53-60 1890 31 1.6 31 1.6 100 11 2 61-77 3524 104 3.0 51 1.4 4912 2 78-91 2737 86 3.1 29 1.1 34 13 2  92-111 4591 119 2.6 58 1.3 49 143 297-313 2808 109 3.9 74 2.6 68 15 2 129-145 2108 117 5.6 44 2.1 38 162 146-162 2467 56 2.3 34 1.4 61 17 2 163-174 2186 57 2.6 23 1.1 40Totals: 34358 1049 3.1 549 1.60 52.3

Table 3, below, lists the 366 selections made on the basis of highthebaine and low oripavine content in latex from leaf samples. Thealkaloid profile is based on peak area, not alkaloid concentration.

TABLE 3 M1 Alkaloid profile (percentage of area under peaks) Seln SeedPlate/ Mor- Co- Lauda- Papav- The- Selection No. Line Tray Positionphine Oripavine Salutaridine deine Reticuline nine erine baine Noscapinename 29 105 7 P20 B10 2 1.7 1 0 0 1 0 94 0 FN1-105-2 41 165 9 P27 B8 04.9 1 0 1 2 0 90 1 FN1-165-1 42 165 9 P27 B9 0 4.4 2 0 2 2 0 90 0FN1-165-2 46 183 10 P29 C6 0 6.5 1 0 1 1 0 91 0 FN1-183-1 54 223 12 P34B1 0 5.7 2 0 0 2 0 91 0 FN1-223-1 64 269 13 P39 D9 0 3.7 1 0 1 1 0 92 0FN1-269-1 65 270 13 P39 E3 0 4.5 2 0 1 1 0 90 0 FN1-270-1 66 272 13 P39G8 0 2.3 1 0 2 1 0 92 0 FN1-272-1 68 291 14 P42 B12 0 2.2 2 0 1 1 0 93 0FN1-291-1 72 300 15 P43 C10 0 6.3 1 0 1 1 0 90 1 FN1-300-2 74 317 15 P45D2 0 2.4 1 0 0 0 0 96 0 FN1-317-1 77 329 16 P46 H2 0 4.9 1 0 1 1 0 92 0FN1-329-1 78 333 16 P47 D10 0 5.4 1 0 1 0 0 93 0 FN1-333-1 79 340 16 P48C1 0 5.8 0 0 1 1 0 91 1 FN1-340-1 80 340 16 P48 C2 0 6.1 0 0 1 1 0 91 1FN1-340-2 83 340 16 P48 C10 1 6.4 1 0 1 1 0 90 0 FN1-340-5 84 345 16 P48H1 0 2.6 1 0 2 1 0 92 1 FN1-345-1 85 345 16 P48 H5 0 4.9 1 0 2 1 0 91 1FN1-345-2 94 399 19 P55 F9 0 1.5 0 0 1 2 0 92 1 FN1-399-3 95 399 19 P55F11 0 2.1 0 0 1 0 3 90 0 FN1-399-4 96 404 19 P56 C5 0 1.5 0 0 1 1 0 93 0FN1-404-1 100 461 21 P63 D6 0 3.6 1 0 1 1 1 92 0 FN1-461-1 103 486 22P66 D1 0 3.5 1 0 2 2 0 91 1 FN1-486-1 106 557 25 P75 C1 0 1.1 0 0 0 0 098 0 FN1-557-1 107 593 27 P79 G5 0 3.9 0 0 1 1 0 94 1 FN1-593-1 110 60127 P80 G6 0 0.4 0 0 1 0 0 97 1 FN1-601-1 111 607 27 P81 E12 0 3.5 0 0 10 0 95 0 FN1-607-1 114 622 28 P83 D9 0 4.3 0 0 1 1 0 93 1 FN1-622-1 115625 28 P83 G12 0 3.6 0 0 1 1 0 94 0 FN1-625-1 117 630 28 P84 D11 0 4.4 00 0 0 0 96 0 FN1-630-1 119 633 28 P84 G9 0 4.3 0 0 2 1 0 92 1 FN1-633-1120 503 23 P68 E8 0 3.3 1 0 1 0 0 94 0 FN1-503-1 122 640 29 P85 F8 0 4.00 0 1 1 0 93 1 FN1-640-1 123 658 29 P87 H4 0 5.8 0 0 2 1 0 90 1FN1-658-1 124 674 30 P89 H7 0 2.8 0 0 1 1 0 95 1 FN1-674-1 125 675 30P90 A10 0 1.5 0 0 1 2 0 94 1 FN1-675-1 127 736 33 P97 F8 0 3.9 0 0 1 1 093 1 FN1-736-1 128 743 33 P98 E8 0 4.2 0 0 3 1 0 91 1 FN1-743-1 129 74833 P99 B7 0 4.8 0 0 2 1 0 91 1 FN1-748-1 130 754 33 P99 H8 0 5.6 0 0 2 10 91 1 FN1-754-1 131 754 33 P99 H12 0 4.1 0 0 1 1 0 93 1 FN1-754-2 132775 34 P102 E8 0 3.9 0 0 2 1 0 93 0 FN1-775-1 134 695 31 P92 E4 0 9.0 00 0 0 0 90 1 FN1-695-2 139 809 36 P106 G1 0 1.7 0 0 0 8 0 91 0 FN1-809-1140 809 36 P106 G2 0 0.0 0 0 0 0 4 96 0 FN1-809-2 143 841 37 P110 G10 42.1 0 0 0 0 0 94 0 FN1-841-1 144 846 37 P111 D1 1 2.0 0 0 1 1 0 95 0FN1-846-1 145 846 37 P111 D2 0 0.0 0 0 0 0 0 100 0 FN1-846-2 146 846 37P111 D12 0 1.1 0 0 2 0 0 97 0 FN1-846-3 147 874 38 P114 H5 0 1.6 0 0 0 00 98 0 FN1-874-1 148 874 38 P114 H6 0 3.2 0 0 0 0 0 97 0 FN1-874-2 149875 39 P115 A5 0 0.0 0 0 0 0 0 100 0 FN1-875-1 150 884 39 P116 B9 3 2.00 0 0 0 0 95 0 FN1-884-1 153 900 40 P118 B1 0 0.0 0 0 0 0 0 98 1FN1-900-1 154 900 40 P118 B3 0 0.5 0 0 0 1 0 97 1 FN1-900-2 155 900 40P118 B6 0 0.3 0 0 0 0 0 97 1 FN1-900-3 156 900 40 P118 B8 0 0.0 0 0 0 00 98 1 FN1-900-4 157 900 40 P118 B12 0 0.0 0 0 1 1 0 98 0 FN1-900-5 158902 40 P118 D3 0 1.6 0 0 0 0 0 97 0 FN1-902-1 159 912 40 P119 F4 0 1.8 00 1 0 1 92 1 FN1-912-1 160 915 40 P120 A6 0 1.9 0 0 0 1 0 93 2 FN1-915-1161 916 40 P120 B12 0 1.2 0 0 0 1 0 98 0 FN1-916-1 162 945 41 P123 G8 02.5 0 0 0 0 0 94 1 FN1-945-1 163 945 41 P123 G11 0 1.6 0 0 0 0 0 96 0FN1-945-2 167 998 44 P130 D6 0 0.8 0 0 0 0 0 99 0 FN1-998-1 168 998 44P130 D8 0 2.1 0 0 0 0 0 97 0 FN1-998-2 172 1027 45 P134 A4 0 0.9 0 0 0 00 98 0 FN1-1027-1 173 1027 45 P134 A10 0 1.4 0 0 0 0 0 98 0 FN1-1027-2174 1050 46 P136 H5 0 1.6 0 0 0 0 0 98 0 FN1-1050-1 175 1050 46 P136 H110 2.5 0 0 0 1 0 96 0 FN1-1050-2 176 1085 47 P141 C5 0 2.7 0 0 0 0 0 96 0FN1-1085-1 178 1108 48 P144 B, 3 0 0.0 3 0 0 0 0 97 0 FN1-1108-1 1791116 49 P145 B, 9 0 3.6 0 0 0 0 0 94 1 FN1-1116-1 180 1123 49 P146 A, 110 3.6 0 0 0 0 0 95 0 FN1-1123-1 181 1133 49 P147 C, 9 0 1.3 0 0 0 1 0 932 FN1-1133-1 183 1139 50 P148 A, 5 0 2.9 0 0 0 0 1 95 1 FN1-1139-1 1841139 50 P148 A, 8 0 3.8 0 1 0 0 2 92 0 FN1-1139-2 185 1141 50 P148 C, 60 0.0 1 1 0 0 0 95 1 FN1-1141-1 187 1149 50 P149 C, 3 0 1.3 0 0 0 0 0 972 FN1-1149-1 188 1153 50 P149 G, 4 0 0.4 0 0 0 0 1 97 0 FN1-1153-1 1921170 51 P151 H, 2 0 3.6 0 0 0 0 0 94 1 FN1-1170-1 194 1176 51 P152 F, 120 3.9 0 0 0 0 0 93 1 FN1-1176-1 195 1180 51 P153 B, 7 0 0.7 0 2 0 1 0 922 FN1-1180-1 196 1180 51 P153 B, 10 0 2.5 0 0 0 0 0 97 0 FN1-1180-2 1971183 51 P153 E, 9 0 0.9 0 0 0 0 2 95 1 FN1-1183-1 198 1183 51 P153 E, 110 0.8 0 0 0 0 0 94 3 FN1-1183-2 199 1185 51 P153 G, 8 0 0.4 0 1 0 0 0 980 FN1-1185-1 200 1186 51 P153 H, 9 0 2.2 0 0 0 0 0 95 1 FN1-1186-1 2031201 52 P155 G, 3 0 3.5 0 0 1 0 0 93 1 FN1-1201-1 204 1204 52 P156 B, 10 3.0 0 0 0 0 0 95 1 FN1-1204-1 205 1211 52 P156 G, 2 0 2.5 0 0 0 0 0 950 FN1-1211-1 208 1242 54 P160 E1 0 0.0 0 0 0 0 0 100 0 FN1-1242-1 2091242 54 P160 E8 0 0.0 0 0 0 0 0 100 0 FN1-1242-2 210 1242 54 P160 E9 00.0 0 0 0 0 0 100 0 FN1-1242-3 213 1270 55 P164 A11 0 0.0 0 0 0 0 0 1000 FN1-1270-1 214 1272 55 P164 C6 0 0.0 0 0 0 0 0 100 0 FN1-1272-1 2161303 56 P168 B, 2 0 4.3 0 0 1 1 0 92 1 FN1-1303-1 217 1313 57 P169 D, 80 5.2 0 0 0 0 1 94 0 FN1-1313-1 218 1326 57 P171 A, 5 0 3.9 0 0 0 0 0 931 FN1-1326-1 219 1326 57 P171 A, 11 0 2.5 0 1 0 0 1 94 1 FN1-1326-2 2201331 57 P171 F, 7 0 2.3 0 0 0 0 0 96 1 FN1-1331-1 224 1366 59 P176 3: A,11 0 1.9 0 0 0 0 0 98 0 FN1-1366-1 225 1370 59 P176 3: E, 6 0 2.2 0 0 00 0 97 0 FN1-1370-1 226 1373 59 P176 3: H, 11 0 0.5 0 0 0 0 0 99 0FN1-1373-1 227 1376 59 P177 4: C, 4 0 0.0 0 0 0 0 0 98 0 FN1-1376-1 2291381 59 P177 4: H, 11 0 2.2 0 0 0 0 0 97 1 FN1-1381-1 230 1387 60 P1785: F, 1 0 1.2 0 0 0 0 0 97 0 FN1-1387-1 232 1401 60 P180 7: D, 11 0 0.00 0 0 0 0 95 1 FN1-1401-1 233 1402 60 P180 7: E, 12 0 0.0 0 0 0 0 0 96 3FN1-1402-1 234 1403 60 P180 7: F, 1 0 1.2 0 0 0 0 0 95 1 FN1-1403-1 2351405 60 P180 7: H, 5 0 0.0 0 0 0 0 0 96 2 FN1-1405-1 236 1405 60 P180 7:H, 6 0 0.0 0 0 0 0 1 97 0 FN1-1405-2 237 1413 61 P181 H4 0 0.0 0 0 0 0 0100 0 FN1-1413-1 238 1419 61 P182 F2 0 0.0 0 0 0 0 0 100 0 FN1-1419-1239 1432 62 P184 C3 0 0.0 0 0 0 0 0 100 0 FN1-1432-1 240 1444 62 P185 G10 0.0 0 0 0 0 0 100 0 FN1-1444-1 241 1447 62 P186 B3 0 0.0 0 0 0 0 0 1000 FN1-1447-1 242 1474 63 P189 E7 0 0.0 0 0 0 0 0 100 0 FN1-1474-1 2451519 65 P195 B6 0 0.0 0 0 0 0 0 100 0 FN1-1519-1 246 1519 65 P195 B7 00.0 0 0 0 0 0 100 0 FN1-1519-2 248 1533 66 P196 H12 0 0.0 0 0 0 0 0 1000 FN1-1533-1 249 1534 66 P197 A5 0 0.0 0 0 0 0 0 100 0 FN1-1534-1 2501534 66 P197 A6 0 0.0 0 0 0 0 0 100 0 FN1-1534-2 251 1535 66 P197 B11 00.0 0 0 0 0 0 100 0 FN1-1535-1 252 1536 66 P197 C7 0 1.7 0 0 0 0 0 98 0FN1-1536-1 254 1567 67 P201 B7 0 0.0 0 0 0 0 0 100 0 FN1-1567-1 255 157167 P201 F10 0 0.0 0 0 0 0 0 100 0 FN1-1571-1 256 1571 67 P201 F11 0 0.00 0 0 0 0 100 0 FN1-1571-2 257 1571 67 P201 F12 0 0.0 0 0 0 2 0 98 0FN1-1571-3 258 1571 67 P201 F2 0 0.0 0 0 0 0 0 100 0 FN1-1571-4 259 157167 P201 F3 0 0.0 0 0 0 0 0 100 0 FN1-1571-5 260 1571 67 P201 F4 0 0.0 00 0 0 0 100 0 FN1-1571-6 261 1571 67 P201 F5 0 0.0 0 0 0 0 0 100 0FN1-1571-7 262 1571 67 P201 F7 0 0.0 0 0 0 0 0 100 0 FN1-1571-8 263 157167 P201 F8 0 0.0 0 0 0 0 0 100 0 FN1-1571-9 264 1571 67 P201 F9 0 0.0 00 0 0 0 100 0 FN1-1571-10 265 1573 67 P201 H12 0 0.0 0 0 0 0 0 100 0FN1-1573-1 268 1600 69 P205 C11 0 0.0 0 0 0 0 0 100 0 FN1-1600-1 2691621 69 P207 H4 0 0.0 0 0 0 0 0 100 0 FN1-1621-1 270 1625 70 P208 D2 01.9 2 0 1 2 0 94 0 FN1-1625-1 271 1659 71 P212 F8 0 1.4 0 0 2 0 0 97 0FN1-1659-1 272 1660 71 P212 G8 0 1.9 0 0 1 0 0 97 0 FN1-1660-1 273 166271 P213 A11 0 1.8 1 0 1 0 0 96 0 FN1-1662-1 274 1662 71 P213 A12 0 1.5 00 1 0 0 97 0 FN1-1662-2 275 1701 73 P217 H2 0 1.0 0 0 0 0 0 99 0FN1-1701-1 276 1702 73 P218 A11 0 2.0 0 0 0 0 0 97 1 FN1-1702-1 277 170373 P218 B2 0 1.3 0 0 0 0 0 99 0 FN1-1703-1 279 1719 74 P220 B1 0 0.0 0 00 0 0 100 0 FN1-1719-1 280 1741 74 P222 H8 0 1.5 0 0 0 0 0 98 0FN1-1741-1 281 1741 74 P222 H12 0 0.0 0 0 0 0 0 100 0 FN1-1741-2 2821744 75 P223 C9 0 0.0 0 0 0 0 0 100 0 FN1-1744-1 283 1763 75 P225 F8 00.0 0 0 0 0 0 100 0 FN1-1763-1 284 1771 76 P226 F2 0 0.8 1 0 0 0 0 98 0FN1-1771-1 285 1771 76 P226 F3 0 0.0 0 0 0 0 0 100 0 FN1-1771-2 287 181377 P231 H11 0 1.8 1 0 1 0 0 94 0 FN1-1813-1 290 1835 78 P234 C9 0 1.9 00 3 3 0 92 0 FN1-1835-2 291 1835 78 P234 C11 0 1.8 0 0 4 2 0 92 0FN1-1835-3 292 1841 79 P235 D8 0 0.0 0 0 0 0 0 100 0 FN1-1841-1 295 186980 P238 H5 0 0.9 0 0 0 0 0 99 0 FN1-1869-1 296 1869 80 P238 H8 0 1.4 0 00 0 0 98 0 FN1-1869-2 298 1898 81 P242 E5 0 1.3 0 0 1 1 0 97 0FN1-1898-1 299 1913 82 P244 D4 0 0.0 0 0 0 0 0 100 0 FN1-1913-1 301 194483 P248 C1 0 1.6 0 0 1 0 0 97 0 FN1-1944-1 304 2017 86 P257 C12 0 1.0 10 0 0 0 98 0 FN1-2017-1 307 2085 89 P265 G1 0 2.3 0 0 0 0 0 96 0FN1-2085-1 308 2085 89 P265 G2 0 1.4 0 0 0 0 0 99 0 FN1-2085-2 309 210389 P267 H2 0 1.2 0 0 0 0 0 99 0 FN1-2103-1 310 2114 90 P269 C2 0 0.8 1 01 0 0 97 0 FN1-2114-1 311 2115 90 P269 D5 0 1.0 0 0 0 0 0 99 0FN1-2115-1 312 2152 91 P273 H3 0 0.8 0 0 0 0 0 99 0 FN1-2152-1 313 215291 P273 H7 0 0.0 0 0 0 0 0 100 0 FN1-2152-2 314 2152 91 P273 H9 0 0.8 00 0 0 0 99 0 FN1-2152-3 315 2152 91 P273 H11 0 0.0 0 0 0 0 0 100 0FN1-2152-4 317 2154 92 P274 B4 0 4.1 0 0 0 0 4 92 0 FN1-2154-1 318 217292 P276 D7 0 0.0 0 0 0 0 0 100 0 FN1-2172-1 319 2175 92 P276 G8 0 0.0 00 0 0 0 100 0 FN1-2175-1 321 2186 93 P278 B6 0 2.0 0 0 0 0 0 98 0FN1-2186-1 322 2186 93 P278 B11 0 0.0 0 0 0 0 0 98 0 FN1-2186-2 324 219693 P279 D5 0 1.3 0 1 1 3 0 94 0 FN1-2196-1 325 2199 93 P279 G3 0 0.0 0 00 0 0 100 0 FN1-2199-1 326 2199 93 P279 G10 0 0.0 0 0 0 0 0 100 0FN1-2199-2 327 2200 93 P279 H1 0 0.0 0 0 0 0 0 97 0 FN1-2200-1 330 221594 P281 G2 0 0.0 0 0 0 0 3 91 0 FN1-2215-1 331 2215 94 P281 G6 0 0.0 0 00 0 0 97 0 FN1-2215-2 332 2219 94 P282 B2 0 0.0 0 0 0 0 0 100 0FN1-2219-1 333 2219 94 P282 B6 0 0.0 0 0 0 0 0 100 0 FN1-2219-2 335 222194 P282 D8 0 0.0 4 0 0 0 4 93 0 FN1-2221-1 336 2224 94 P282 G4 0 0.0 0 00 0 0 100 0 FN1-2224-1 337 2231 95 P283 F5 0 0.0 0 0 0 0 0 100 0FN1-2231-1 338 2231 95 P283 F12 0 0.0 0 0 0 0 0 100 0 FN1-2231-2 3392233 95 P283 H7 0 3.0 0 0 0 0 0 97 0 FN1-2233-1 340 2241 95 P284 H1 00.0 0 0 0 0 0 100 0 FN1-2241-1 341 2241 95 P284 H4 0 2.7 0 0 0 0 0 97 0FN1-2241-2 342 2243 95 P285 B9 0 0.0 0 0 0 0 0 96 0 FN1-2243-1 343 224595 P285 C5 0 0.0 0 0 0 0 0 98 0 FN1-2245-1 344 2245 95 P285 C10 0 0.0 00 0 0 0 100 0 FN1-2245-2 345 2255 96 P286 E9 0 0.0 0 0 0 0 0 100 0FN1-2255-1 346 2267 96 P288 A10 1 0.0 1 0 0 0 0 99 0 FN1-2267-1 349 228097 P289 F11 0 0.0 0 0 0 5 0 91 4 FN1-2280-1 354 2288 97 P290 F9 0 0.9 00 1 1 0 96 1 FN1-2288-1 355 2325 99 P295 C5 0 0.0 0 0 0 0 0 100 0FN1-2325-1 356 2329 99 P295 G4 0 0.0 0 0 2 2 0 97 0 FN1-2329-1 357 2365100 P300 C9 0 0.0 0 0 0 2 0 98 0 FN1-2365-1 358 2372 101 P301 B1 0 0.0 00 0 0 0 100 0 FN1-2372-1 361 2412 102 P306 B2 0 0.7 0 0 0 0 0 99 0FN1-2412-1 362 2425 103 P307 G8 0 0.0 0 0 0 0 0 100 0 FN1-2425-1 3632427 103 P308 A6 0 0.0 0 0 0 0 0 100 0 FN1-2427-1 364 2437 103 P309 C120 0.0 0 0 0 0 0 100 0 FN1-2437-1 365 2443 104 P310 A1 0 0.7 0 0 1 0 0 970 FN1-2443-1 367 2492 106 P316 A5 0 0.0 0 0 0 1 0 99 0 FN1-2492-1 3742604 110 P330 A6 0 0.0 0 0 0 1 0 99 0 FN1-2604-1 376 7299 298 P892 C2 00.0 0 0 0 3 0 97 0 FN1-7299-2 377 7299 298 P892 C3 0 0.0 0 0 0 2 0 98 0FN1-7299-3 378 7299 298 P892 C4 0 0.5 0 0 0 1 0 98 0 FN1-7299-4 379 7299298 P892 C10 0 0.7 0 0 0 2 0 98 0 FN1-7299-5 380 7299 298 P892 C12 0 0.00 0 0 3 0 97 0 FN1-7299-6 381 7303 298 P892 G10 0 0.0 0 0 0 0 0 100 0FN1-7303-1 382 7304 298 P892 H2 0 0.0 0 0 0 0 0 100 0 FN1-7304-1 3837304 298 P892 H5 0 0.0 0 0 1 1 0 98 0 FN1-7304-2 384 7306 298 P893 B6 00.0 0 0 0 1 0 98 1 FN1-7306-1 385 7325 299 P895 E10 0 0.0 1 0 1 2 0 97 0FN1-7325-1 386 7329 299 P896 A8 0 0.0 0 0 0 0 0 100 0 FN1-7329-1 3877329 299 P896 A10 0 0.0 0 0 0 1 0 99 0 FN1-7329-2 388 7332 299 P896 D3 00.0 0 0 0 1 0 99 0 FN1-7332-1 389 7341 299 P897 E3 0 0.0 0 0 0 0 0 100 0FN1-7341-1 390 7342 299 P897 F1 0 0.0 0 1 0 2 0 97 0 FN1-7342-1 391 7342299 P897 F9 0 0.0 0 1 0 2 0 96 1 FN1-7342-2 393 7348 300 P898 D5 0 1.1 00 0 0 0 99 0 FN1-7348-1 394 7353 300 P898 H1 0 0.6 0 0 0 0 0 99 0FN1-7353-1 395 7354 300 P899 A3 0 0.0 0 0 0 1 0 99 0 FN1-7354-1 396 7354300 P899 A8 0 0.0 0 0 0 1 0 99 0 FN1-7354-2 397 7355 300 P899 B2 0 0.0 00 0 0 0 100 0 FN1-7355-1 398 7355 300 P899 B5 0 0.0 0 0 0 1 0 99 0FN1-7355-2 399 7373 301 P901 C10 0 1.0 0 0 0 0 0 99 0 FN1-7373-1 4007378 301 P901 H1 0 0.0 0 0 1 0 0 99 0 FN1-7378-1 401 7380 301 P902 B9 00.0 0 0 0 0 0 100 0 FN1-7380-1 402 7386 301 P902 H6 0 0.0 0 0 1 1 0 98 0FN1-7386-1 403 7386 301 P902 H9 0 0.0 0 0 0 0 0 100 0 FN1-7386-2 4047397 302 P904 B1 0 0.0 0 0 0 0 0 100 0 FN1-7397-1 405 7399 302 P904 D3 00.0 0 0 0 3 0 97 0 FN1-7399-1 406 7400 302 P904 E12 0 0.0 0 0 0 0 0 1000 FN1-7400-1 407 7445 303 P909 H12 0 0.9 0 0 0 1 0 98 0 FN1-7445-1 4087447 304 P910 B9 0 0.7 0 0 0 0 0 98 0 FN1-7447-1 409 7448 304 P910 C6 00.0 0 0 0 0 0 98 2 FN1-7448-1 410 7452 304 P910 G1 0 0.0 0 0 1 1 0 98 0FN1-7452-1 411 7462 304 P911 H6 0 0.9 0 0 0 0 1 99 0 FN1-7462-1 412 7467304 P912 C1 0 0.6 0 0 0 1 0 97 0 FN1-7467-1 413 7467 304 P912 C2 0 0.4 10 0 2 0 97 0 FN1-7467-2 414 7471 304 P912 G7 0 1.0 0 0 0 0 0 98 1FN1-7471-1 415 7472 304 P912 H2 0 0.0 0 0 0 0 0 95 5 FN1-7472-1 416 7491305 P914 G8 0 0.8 0 0 0 0 0 99 0 FN1-7491-1 417 7499 305 P915 F2 0 1.2 00 0 0 0 99 0 FN1-7499-1 418 7506 306 P916 B6 0 1.1 0 0 1 0 1 97 0FN1-7506-1 419 7509 306 P916 D1 0 0.0 0 0 0 0 0 100 0 FN1-7509-1 4207513 306 P916 H5 0 1.0 0 0 1 1 0 96 1 FN1-7513-1 421 7525 306 P918 C10 01.3 0 0 1 1 0 95 1 FN1-7525-1 422 7529 306 P918 G11 0 0.0 0 0 0 0 0 1000 FN1-7529-1 423 7535 307 P919 E8 0 0.0 0 0 0 0 0 97 3 FN1-7535-1 4247536 307 P919 F1 0 1.2 0 0 2 1 0 96 0 FN1-7536-1 426 7551 307 P921 C5 00.0 0 0 0 0 0 100 0 FN1-7551-1 427 7557 308 P922 A12 0 0.0 0 0 0 0 0 1000 FN1-7557-1 428 7558 308 P922 B7 0 0.0 0 0 0 0 0 100 0 FN1-7558-1 4297560 308 P922 D8 0 1.2 0 0 0 1 0 98 0 FN1-7560-1 430 7566 308 P923 B9 00.0 0 0 0 0 2 98 0 FN1-7566-1 431 7579 308 P924 F3 0 0.8 0 0 3 0 2 92 2FN1-7579-1 432 7584 309 P925 B3 0 0.0 0 0 1 1 0 98 0 FN1-7584-1 433 7587309 P925 E7 0 0.0 0 0 0 0 0 100 0 FN1-7587-1 434 7592 309 P926 B2 0 0.00 0 0 0 0 100 0 FN1-7592-1 435 7592 309 P926 B4 0 0.7 0 0 1 1 0 97 0FN1-7592-2 436 7598 309 P926 H3 0 0.7 0 0 1 2 0 95 1 FN1-7598-1 437 7598309 P926 H10 0 0.7 1 0 1 1 0 96 1 FN1-7598-2 438 7600 309 P927 B1 0 0.00 0 0 0 0 100 0 FN1-7600-1 439 7600 309 P927 B5 0 0.0 0 0 0 0 0 100 0FN1-7600-2 440 7629 310 P930 C8 0 1.1 0 1 0 0 0 95 1 FN1-7629-1 441 7640311 P931 B4 0 0.0 0 1 0 0 0 96 1 FN1-7640-1 442 7647 311 P931 F1 0 0.0 03 0 0 0 96 0 FN1-7647-1 443 7656 311 P932 F3 0 1.7 0 1 0 0 0 94 1FN1-7656-1 444 7666 311 P933 G8 0 1.0 0 1 0 0 0 95 1 FN1-7666-1 445 7674312 P934 F3 0 1.4 0 1 0 0 0 96 1 FN1-7674-1 446 7686 312 P936 A6 0 0.0 00 0 0 0 100 0 FN1-7686-1 447 7702 313 P937 H8 0 0.9 0 0 0 0 0 98 1FN1-7702-1 448 7718 313 P939 G6 0 0.0 0 0 0 1 0 99 0 FN1-7718-1 450 3122132 P394 E2 0 0.0 0 0 0 5 0 95 0 FN1-3122-1 451 3123 132 P394 F12 0 0.00 0 0 5 0 95 0 FN1-3123-1 452 3132 132 P395 G11 0 0.0 0 0 0 6 0 94 0FN1-3132-1 453 3141 132 P396 G1 0 0.8 0 0 1 1 0 96 1 FN1-3141-1 454 3141132 P396 G10 0 0.0 0 0 0 0 4 96 0 FN1-3141-2 455 3158 133 P398 G6 0 0.00 0 0 3 0 97 0 FN1-3158-1 456 3176 134 P400 G1 0 0.9 0 0 3 1 0 95 0FN1-3176-1 457 3206 135 P404 E3 0 0.0 0 0 0 0 4 96 0 FN1-3206-1 458 3209135 P404 H11 0 0.0 0 0 1 0 2 96 1 FN1-3209-1 459 3215 135 P405 F5 0 1.00 0 1 0 2 95 1 FN1-3215-1 460 3228 136 P407 C8 0 0.0 0 0 1 2 0 97 0FN1-3228-1 461 3258 137 P410 G4 0 0.0 0 0 1 0 4 94 1 FN1-3258-1 462 3270138 P412 B3 0 0.0 0 0 0 5 0 95 0 FN1-3270-1 463 3288 138 P414 C7 0 0.0 00 0 0 3 97 0 FN1-3288-1 464 3295 139 P415 A2 0 0.0 0 0 2 4 0 94 0FN1-3295-1 465 3296 139 P415 B10 0 0.0 0 0 0 3 0 97 0 FN1-3296-1 4663296 139 P415 B12 0 1.0 0 0 0 2 0 97 0 FN1-3296-2 467 3297 139 P415 C120 0.0 0 0 1 0 3 95 0 FN1-3297-1 468 3299 139 P415 E1 0 0.0 0 0 0 0 3 961 FN1-3299-1 469 3300 139 P415 F2 0 0.0 0 0 1 0 3 95 0 FN1-3300-1 4723310 139 P416 H4 0 0.0 0 0 0 2 0 96 1 FN1-3310-1 473 3320 140 P418 B10 00.6 0 0 1 2 0 96 1 FN1-3320-1 474 3326 140 P418 H2 0 0.0 0 0 0 2 0 98 0FN1-3326-1 475 3328 140 P419 B2 0 0.0 0 0 1 1 0 98 1 FN1-3328-1 476 3328140 P419 B7 0 0.0 0 0 0 0 7 91 0 FN1-3328-2 477 3306 139 P416 D7 0 3.9 00 0 0 1 94 1 FN1-3306-1 478 3365 141 P423 F3 0 0.0 0 0 0 7 0 93 0FN1-3365-1 479 3368 141 P423 H10 0 0.0 0 0 0 0 0 100 0 FN1-3368-1 4803376 142 P424 H9 0 0.0 0 0 0 2 0 98 0 FN1-3376-1 481 3376 142 P424 H11 00.0 0 0 0 2 0 98 0 FN1-3376-2 482 3383 142 P425 G9 0 0.0 0 0 0 0 0 100 0FN1-3383-1 483 3386 142 P426 B9 0 0.0 0 0 0 2 0 98 0 FN1-3386-1 484 3387142 P426 C5 0 0.0 0 0 1 1 0 98 0 FN1-3387-1 485 3387 142 P426 C7 0 0.0 00 0 0 0 98 2 FN1-3387-2 486 3388 142 P426 D7 0 0.0 0 0 0 1 0 97 1FN1-3388-1 487 3406 143 P428 F12 0 0.0 0 0 0 0 0 100 0 FN1-3406-1 4883408 143 P428 H11 0 1.0 0 0 1 1 0 97 0 FN1-3408-1 489 3413 143 P429 E2 00.0 0 0 2 0 0 98 0 FN1-3413-1 491 3444 145 P433 C12 0 0.0 0 0 0 0 0 1000 FN1-3444-1 493 3488 146 P438 F4 0 0.0 0 0 2 0 0 98 0 FN1-3488-1 4943492 147 P439 B4 0 0.0 0 0 4 2 0 93 0 FN1-3492-1 495 3497 147 P439 G3 00.0 0 0 0 0 0 100 0 FN1-3497-1 496 3497 147 P439 G4 0 0.0 0 0 0 0 0 1000 FN1-3497-2 497 3497 147 P439 G11 0 0.0 0 0 0 0 0 100 0 FN1-3497-3 4983531 148 P443 H1 0 0.0 0 0 0 0 0 100 0 FN1-3531-1 500 3608 151 P453 C8 01.1 0 0 1 0 1 97 0 FN1-3608-1 501 3612 151 P453 G6 0 0.0 0 0 0 0 0 100 0FN1-3612-1 503 3635 152 P456 F1 0 1.1 0 0 1 1 0 96 1 FN1-3635-1 504 3635152 P456 F2 0 0.0 0 0 0 0 0 98 0 FN1-3635-2 505 3635 152 P456 F4 0 0.7 00 0 0 0 95 0 FN1-3635-3 506 3635 152 P456 F9 0 0.5 0 0 0 0 0 99 0FN1-3635-4 507 3635 152 P456 F10 0 0.0 0 0 0 0 0 100 0 FN1-3635-5 5083635 152 P456 F11 0 0.2 0 0 0 0 0 99 0 FN1-3635-6 509 3679 154 P461 G5 00.6 0 0 0 1 0 98 0 FN1-3679-1 510 3708 155 P465 D2 0 0.0 0 0 0 0 0 100 0FN1-3708-1 511 3710 155 P465 F5 0 0.0 0 0 0 0 0 100 0 FN1-3710-1 5123718 156 P466 F5 0 0.0 0 0 0 0 0 100 0 FN1-3718-1 513 3718 156 P466 F7 01.0 0 0 0 0 0 98 1 FN1-3718-2 514 3770 158 P473 B8 0 1.6 0 0 1 0 0 97 0FN1-3770-1 516 3794 159 P476 A7 0 0.4 0 0 0 0 0 100 0 FN1-3794-1 5173799 159 P476 F4 0 0.9 0 0 1 0 0 97 1 FN1-3799-1 518 3803 159 P477 B9 00.9 0 0 1 0 0 98 1 FN1-3803-1 519 3804 159 P477 C6 0 1.0 0 0 1 0 0 96 1FN1-3804-1 520 3805 159 P477 D12 0 1.0 0 0 1 0 0 98 1 FN1-3805-1 5213817 160 P478 H11 0 0.0 0 0 0 0 0 100 0 FN1-3817-1 522 3821 160 P479 D10 1.1 0 0 1 0 0 98 0 FN1-3821-1 523 3821 160 P479 D3 0 1.4 0 0 0 0 0 981 FN1-3821-2 524 3821 160 P479 D10 0 0.0 0 0 0 0 0 100 0 FN1-3821-3 5253827 160 P480 B6 0 0.7 0 0 0 0 0 99 0 FN1-3827-1 526 3841 161 P481 H3 00.0 0 0 3 0 0 96 1 FN1-3841-1 528 3978 166 P498 H2 0 0.0 0 0 0 0 0 100 0FN1-3978-1 529 3995 167 P501 A5 0 0.0 0 0 0 0 0 100 0 FN1-3995-1 5314027 169 P505 A6 0 0.0 0 0 0 0 0 100 0 FN1-4027-1 532 4027 169 P505 A8 00.6 0 0 0 0 0 99 0 FN1-4027-2 534 4050 169 P507 H3 0 0.4 0 0 1 0 0 99 0FN1-4050-1 535 4053 170 P508 C9 0 0.7 0 0 0 0 0 99 0 FN1-4053-1 537 4093171 P513 C5 0 1.0 0 0 2 1 0 94 1 FN1-4093-1 541 4121 172 P516 G9 0 1.2 00 0 0 0 97 1 FN1-4121-1 542 4124 173 P517 B2 0 0.7 0 0 1 1 0 97 1FN1-4124-1 543 4128 173 P517 F6 1 1.3 0 0 1 0 0 94 0 FN1-4128-1 546 4144173 P519 F12 0 0.9 0 0 0 0 0 97 0 FN1-4144-1 547 4145 173 P519 G7 0 0.70 0 0 0 0 99 0 FN1-4145-1 548 4145 173 P519 G11 0 0.5 0 0 0 0 0 97 0FN1-4145-2 549 4148 174 P520 B7 0 0.7 0 0 1 1 0 98 0 FN1-4148-1Testing of Poppy Straw Alkaloid Content in M2 Generation

Capsules were harvested from the greenhouse as they matured. Seed wasremoved and weighed into seed envelopes. The poppy straw was placed into50 mL BD Falcon™ tubes (BD Biosciences, San Jose, Calif.) withoutgrinding and dried either on the lab bench for several days at roomtemperature or in the laboratory oven at 50° C. for 3 hours. Wherecapsules were large, only a portion of the capsule was used foranalysis, the rest being discarded.

For analysis, the poppy straw was weighed, and either 5 mL or 10 mL acidextractant (5% ethanol (“EtOH”), 0.17% phosphoric acid) added, dependingon whether the straw samples weighed less or more than 0.2 grespectively. The samples were agitated with a Ratek orbital shaker(Ratek Instruments, Boronia, Victoria, Australia) for 3 hours. Theliquid phase was then filtered using Pall AcroPrep™ 96 filter plates (PNS5045), and the filtrate was analysed for alkaloids using a WatersAcquity UPLC® system (Waters Corporation, Milford, Mass.). The UPLCmethod used was the same 2.5 minute method as used for leaf samples.Additional extractant was transferred to 1.2 mL wells in 96 well plates,sealed and frozen in case further analysis was required.

The alkaloid contents and profiles were calculated from the UPLC resultsand weight data. Where the weight was <0.1 g, the weight was deemed as0.1 g.

The seeds were allowed to dry on the laboratory bench, catalogued andstored.

Of the 549 M2 plants originally selected, 434 survived to produce atleast one capsule for harvest (79%). A further 6 mislabeled capsuleswere harvested, so their M1 parents were not known. 395 plants producedsome seed, although 58 had <0.06 g seed, and only 171 plants producedmore than 1 g seed.

Due to the potential lack of accuracy with the alkaloid content data(due to low capsule weights and large particle sizes), this data wasanalysed further only after conversion to alkaloid profiles: i.e. thealkaloid contents in comparison with the total alkaloid content.

Where multiple capsules were harvested from one plant, the mean wasdetermined using Minitab 14 statistical software (Minitab Inc., StateCollege, Pa.).

Table 4, below, shows the results for 133 M2 selections from 92independent M1 plants that were high in thebaine (>90% of alkaloids) andlow in oripavine (<10%). Thirty six M2 selections from 27 independent M1plants had more than 95% of the alkaloid in the straw as thebaine.Twenty one selections from 16 independent M1 plants had more than 96% ofthe alkaloid in the straw as thebaine. Eight selections from 6independent M1 plants had more than 97% of the alkaloid in the straw asthebaine. One selection, FN1-900-5, was identified that had more than98% of its alkaloid present as thebaine. It can be seen that theoripavine content in the straw of these plants was very low, withseveral selections having less than 1% of the alkaloid combination, andsome with less than 0.5% of the alkaloid combination. All“thebaine-only” plants however contained a small proportion of oripavinein their poppy straw.

TABLE 4 Alkaloid profiles (based on alkaloid concentrations in poppystraw) of M2 plants selected for high thebaine and low oripavine. Meansare shown where the number of capsules (caps) was more than one. Selnno. Seln name Caps Morphine Oripavine Codeine Salutaridine ReticulineLaudanine Papaverine Thebaine Noscapine 68 FN1-291-1 15 0.4 4.7 0 0.20.9 0.1 0.2 93.3 0.2 106 FN1-557-1 1 0 2.8 0 0.3 6.2 0 0 90.7 0 110FN1-601-1 1 0.1 0.4 0 0 1.5 0.2 0 97.6 0.1 139 FN1-809-1 5 0 1.6 0.1 0.64.9 0 0 92.7 0 140 FN1-809-2 2 0 1.7 0.1 0.3 2.1 0 0 95.6 0.2 144FN1-846-1 1 0 0.1 0 0 2.5 0.1 0 97.2 0 145 FN1-846-2 3 2.6 0.4 0 0 3.10.2 0 93.5 0.1 149 FN1-875-1 3 1.8 3.6 0 0.1 1.9 0.1 0 92.4 0.1 153FN1-900-1 3 0 0.6 0 0.1 1.5 0.2 0 97.5 0.1 154 FN1-900-2 3 0 0.6 0 0.21.8 0.2 0 97.1 0.1 155 FN1-900-3 3 0.8 0.5 0 0 2.3 0.2 0 96.1 0.1 156FN1-900-4 7 2.3 0.6 0 0.2 1.5 0.2 0 94.9 0.3 157 FN1-900-5 1 0 0.6 0 0.21 0.1 0 98 0.1 161 FN1-916-1 4 0.6 5.5 0 0.1 1.3 0.1 0 92.3 0.1 167FN1-998-1 1 1.5 1.1 0.6 0 2.4 0 0 91.4 3.1 168 FN1-998-2 2 0 1 0.1 0.32.4 0 0 96.3 0 172 FN1-1027-1 1 0 2.2 0 0 3.1 0 0 94.7 0 173 FN1-1027-24 0.3 2.6 0 0.3 2 0.1 0 94.6 0.2 174 FN1-1050-1 8 3.5 1 0 0.4 1.3 0 093.7 0 175 FN1-1050-2 2 0.2 3 0 0.1 1.7 0 0 94.8 0.2 180 FN1-1123-1 2 01 0 0.4 3.6 0 0 95.2 0 184 FN1-1139-2 1 0 5.7 0 0.1 1.3 0.2 0 92.5 0.1188 FN1-1153-1 10 0.1 7.9 0 0 1.3 0.1 0 90.6 0 199 FN1-1185-1 3 0 2.9 00.1 2.7 0.2 0 93.9 0.2 208 FN1-1242-1 3 1.2 0.6 0 0.1 2.8 0.2 0 95 0.1209 FN1-1242-2 2 0 0.3 0 0 2.8 0.2 0 96.7 0.1 210 FN1-1242-3 3 0 0.4 00.1 1.7 0.1 0 97.5 0.1 219 FN1-1326-2 4 0.5 6.9 0 0.1 1.8 0.1 0 90.6 0.1240 FN1-1444-1 4 0 5.2 0 0 1.4 0.2 0 93.2 0.2 249 FN1-1534-1 6 0.1 0.7 00.5 2.6 0 0 96.1 0 250 FN1-1534-2 2 0.5 1.5 0 0.5 4.9 0 0 92.5 0.2 255FN1-1571-1 2 0.4 1.5 0 0.5 5.7 0 0 92 0 256 FN1-1571-2 2 0.5 1.8 0 0.74.2 0 0 93 0 257 FN1-1571-3 3 0.6 1.5 0 0.5 2.5 0 0 94.7 0 258FN1-1571-4 2 0.7 1.9 0 0.4 6.9 0.1 0 90.2 0 259 FN1-1571-5 1 0 1.5 0 0.73.4 0 0 94.3 0 260 FN1-1571-6 4 0.1 1.2 0.1 0.5 4.9 0 0 93.2 0 261FN1-1571-7 3 0 1.3 0 0.3 5 0 0 93.3 0 262 FN1-1571-8 1 1 1.7 0 0.8 2.3 00 94 0.2 263 FN1-1571-9 3 0.6 1.8 0 0.8 4.6 0 0.1 92.1 0 264 FN1-1571-104 0.2 1.4 0.1 0.4 5 0 0 92.9 0 270 FN1-1625-1 1 0 2.3 0 0.3 1.2 0.3 095.7 0.1 273 FN1-1662-1 1 0 4.6 0 0.1 1.4 0 0.6 93.1 0.3 275 FN1-1701-12 0 2.9 0.1 0.1 2.2 0.2 0 94.6 0.1 279 FN1-1719-1 4 0.2 0.3 0 0 2.6 0.10 96.6 0.1 280 FN1-1741-1 1 0 3.1 0 0 3.4 0.2 0 93 0.3 284 FN1-1771-1 30 2 0 0.3 1.1 0.1 0 96.6 0.1 285 FN1-1771-2 2 0 4.8 0 0.2 1.6 0 0 93.40.1 292 FN1-1841-1 1 0 4.7 0 0 1.3 0.3 0 93.7 0 296 FN1-1869-2 1 0 2.2 00.4 1.7 0.1 0 95.5 0.1 299 FN1-1913-1 3 0 3 0 0 1.8 0 0 95.1 0 307FN1-2085-1 1 0 4 0 0 2.8 0.1 0 93 0.1 309 FN1-2103-1 5 0.1 6.8 0 0.1 2.10.2 0 90.7 0 312 FN1-2152-1 3 0 2.1 0 0.1 5.6 0 0 92.3 0 313 FN1-2152-22 0 0.9 0.2 0.6 3.6 0.1 0 94.8 0 314 FN1-2152-3 3 0 1.1 0 0.7 3.4 0 0.194.7 0 315 FN1-2152-4 2 0 1.2 0 0.5 3.3 0 0 95 0 318 FN1-2172-1 1 0 4 00 4.9 0.2 0 90.8 0.1 319 FN1-2175-1 1 0 8 0 0 1.2 0.1 0 90.4 0.3 321FN1-2186-1 2 0 3.5 0.1 0.4 1.6 0 0.3 94.1 0.1 325 FN1-2199-1 3 0 0.4 00.1 4.3 0.2 0 95 0 326 FN1-2199-2 2 0 0.3 0 0.1 3.6 0.1 0 95.9 0.2 339FN1-2233-1 1 0 2.8 0 0.1 1.4 0 0 95.7 0 343 FN1-2245-1 2 0 6.1 0 0.1 1.50.1 0 92.1 0.2 364 FN1-2437-1 3 0 0.5 0 0.1 3.1 0.2 0 96.2 0 367FN1-2492-1 1 0 5.5 0 0.1 2.2 0.1 0 92.1 0.1 374 FN1-2604-1 3 0 0.3 0.10.1 5.4 0.2 0 93.7 0.2 377 FN1-7299-3 2 0 2.4 0.1 0.2 1.3 0.1 0 95.8 0.2378 FN1-7299-4 2 0 3.3 0.1 0.2 1.7 0.2 0 94.6 0.2 379 FN1-7299-5 1 0 2.70.1 0.4 1.6 0.2 0 94.7 0.3 380 FN1-7299-6 2 0 2 0 0.1 1 0.1 0 96.8 0.1384 FN1-7306-1 1 0 6.8 0 0 0 0 0 93.1 0.1 396 FN1-7354-2 1 0 0.6 0 1.24.3 0 0 93.9 0 400 FN1-7378-1 1 0 4.4 0 0.1 1.7 0 0 93.5 0.3 404FN1-7397-1 1 0 8 0 0 1 0 0 91 0 407 FN1-7445-1 1 0 5.6 0 0 2.3 0.2 091.9 0 408 FN1-7447-1 1 0 3.3 0 0.1 3.5 0.1 0 92.8 0.2 409 FN1-7448-1 10 5.2 0 0.1 2.7 0.1 0 91.8 0.2 414 FN1-7471-1 1 0 3.7 0.1 0 3.2 0.2 092.3 0.4 416 FN1-7491-1 1 0 6.9 0 0.1 1.1 0.2 0 91.8 0 417 FN1-7499-1 10 5.4 0 0 1.6 0.2 0 92.6 0.3 421 FN1-7525-1 2 0 5.5 0 0.1 1.6 0 0.4 92.50.1 422 FN1-7529-1 1 0 6.7 0 0.1 2.2 0 0.9 90 0.2 426 FN1-7551-1 1 0 4.60 0.1 2.3 0.1 0 92.8 0.1 433 FN1-7587-1 1 0 8.4 0 0 0.6 0.1 0 90.8 0 434FN1-7592-1 1 0.5 2 0 0.6 0.5 0 0 96.3 0 436 FN1-7598-1 1 0 5.7 0 0.1 2.20.1 0 91.7 0.2 437 FN1-7598-2 1 0 4.5 0 0 1.8 0 0 93.5 0.3 443FN1-7656-1 1 0 7.7 0 0.1 1.4 0 0.5 90.2 0.2 445 FN1-7674-1 5 0 2.4 0 02.6 0.1 0.2 94.5 0.2 446 FN1-7686-1 2 0 1.5 0 0.4 4.9 0.1 0 93.3 0 447FN1-7702-1 1 0 3.2 0 0.1 2 0 0 94.6 0 453 FN1-3141-1 1 0 4.6 0 0.1 2.30.2 0 92.7 0.1 454 FN1-3141-2 1 0 8.1 0 0 1.6 0.2 0 90.1 0 455FN1-3158-1 2 0 8 0 0.1 1.6 0.2 0 90.2 0.1 458 FN1-3209-1 1 0 5 0 0 2 0.20 92.5 0.2 461 FN1-3258-1 1 0.3 6.1 0 0 1.8 0.2 0 91.6 0 463 FN1-3288-11 0 5.8 0 0.1 1.8 0.2 0 92.1 0.1 465 FN1-3296-1 1 0 8.7 0 0.1 1.1 0.1 090.1 0 466 FN1-3296-2 1 0 2.2 0.4 0.2 1.8 0.2 0 95.2 0.1 474 FN1-3326-11 0 7.2 0 0.1 1.6 0.2 0 90.9 0.1 475 FN1-3328-1 2 0 0.4 0 0.2 6.5 0.3 092.7 0 476 FN1-3328-2 1 0 0.4 0 0.1 2.9 0.2 0 96.3 0.1 477 FN1-3306-1 20 2.8 0.1 0.1 2.5 0.1 0 94.3 0.3 480 FN1-3376-1 1 0 5 0 0 1.4 0.3 0 93.30 483 FN1-3386-1 1 0 4.6 0.1 0.1 2.6 0 1.1 91.2 0.4 486 FN1-3388-1 1 05.5 0 0.1 3.1 0.2 0 90.8 0.3 489 FN1-3413-1 1 0 6.3 0 0.1 2.8 0.1 0 90.60.2 495 FN1-3497-1 1 0 1.4 0 0 4 0 0 94.6 0 496 FN1-3497-2 2 0 1.2 0 0.22.8 0 0 96 0 497 FN1-3497-3 2 0 1.2 0 0.2 4.7 0 0 94 0 503 FN1-3635-1 20 1.2 0 0.3 4 0.1 0 94.3 0.2 504 FN1-3635-2 1 0 1.3 0 0 2.8 0 0 95.9 0506 FN1-3635-4 1 0 1.2 0 0.4 3.6 0 0 94.8 0 507 FN1-3635-5 1 0 1 0 0 1.90 0 97.1 0 508 FN1-3635-6 1 0 1.3 0 0 2.2 0 0 96.5 0 509 FN1-3679-1 1 03.2 0.2 0.1 4.6 0 0 91.7 0.3 511 FN1-3710-1 1 0 8.1 0 0 1.3 0.2 0 90.5 0512 FN1-3718-1 1 0 4 0 0.1 1.5 0.2 0 94.2 0.1 516 FN1-3794-1 2 0 0.1 00.1 4 0.2 0 95.7 0.1 521 FN1-3817-1 1 0 7 0 0 1.5 0.1 0 91.4 0 522FN1-3821-1 1 0.9 5.9 0 0 2.5 0.2 0 90.3 0.3 523 FN1-3821-2 1 0 3.3 0 0.13.9 0.2 0 92 0.4 524 FN1-3821-3 1 0 6.7 0 0 2.3 0.1 0 90.9 0 525FN1-3827-1 1 0 4.7 0 0 1 0.2 0 94.1 0 528 FN1-3978-1 1 0 3 0 0 2.7 0.3 094 0 529 FN1-3995-1 2 0 0.8 0 0.1 3.4 0.2 0 95.5 0.2 534 FN1-4050-1 2 00.1 0 0.1 2.7 0.1 0 97 0.1 535 FN1-4053-1 3 0 5.1 0 0 1.7 0.2 0 93.1 0541 FN1-4121-1 1 0 2.2 0 0 1.6 0.3 0 96 0 542 FN1-4124-1 2 0 6.9 0 0 1.40.1 0 91.8 0 546 FN1-4144-1 1 0 4 0 0 2.9 0.3 0 92.7 0 548 FN1-4145-2 10 1.9 0 0.5 4.1 0 0 93.5 0Growing and Evaluation of M3 Generation

Two of the highest thebaine lines were selected for increase in agreenhouse over winter 2007 to provide data to confirm their alkaloidcomposition and genetic stability. Thus, the plants grown in thisexperiment were the M3 generation.

The pots were sown in the greenhouse on 11 Apr. 2007 in double rows,with 120 pots in each double row. Each pot was thinned to 6 plants.Greenhouse conditions were as used previously except that high intensitylights were used to maintain light intensities of approximately 9900 luxfor 12 hours per day.

At green capsule stage, latex samples were taken from 24 randomly chosenplants from each line. The samples were obtained from the stigmaticdiscs using the ray-pluck technique. A stigmatic ray was removed fromeach plant and dropped into acid extraction solution (5% EtOH, 0.17%H₃PO₄) in a filter plate (Pall AcroPrep™ 96 Filter Plate 0.2 μm GHP,NTRL, 350 μL). The rest of the procedure was the same as for leaf latextests. A separate trial established that there was no significantdifference in thebaine or oripavine results attributable to using acidextraction solution instead of latex extraction solution as usedpreviously.

Table 5 shows the results of the latex testing at the green capsulestage for the M3 Generation. The number of plants randomly sampled andtested is shown as “N”. All plants tested had the same alkaloid profileof high thebaine and substantially no oripavine or morphine.

TABLE 5 Results of the green capsule stage testing. The table shows thepercentage of area under chromatogram peaks. Thebaine Oripavine MorphineCodeine Papaverine Noscapine Thebaine ratio Norman ratio Selection NMean SE Mean SE Mean SE Mean SE Mean SE Mean SE Mean SE Mean SEFN1-900-1 23 98.0 0.1 0.5 0.1 0.2 0.0 0.7 0.0 0.6 0.0 0.0 0.0 0.9950.001 0.99 0.00 FN1-1242-3 24 98.6 0.0 0.2 0.0 0.1 0.0 0.6 0.0 0.5 0.00.0 0.0 0.998 0.000 0.99 0.00 Thebaine ratio is the thebaine areadivided by the total of the thebaine and oripavine areas Norman ratio isthe sum of the thebaine and oripavine areas divided by the sum ofmorphine, oripavine, codeine and thebaine areas

When the capsules were dry, the plants were harvested by hand. Theharvested capsules were weighed, and then threshed and sieved toseparate seed and straw. The straw was sub-sampled and ground to 2 mm.

The straw was extracted using acid extraction solution (5% EtOH, 0.17%H₃PO₄), and analysed using Waters Acquity ULPC® for alkaloid contentagainst standard alkaloid solutions on a dry weight basis. The loss ondrying (LOD) of the straw was determined by heating a sample at 88° C.for 9 minutes using an infrared (IR) balance (A&D Company Ltd ModelAD4717, Japan).

Peak area data was used to calculate alkaloid concentration in the strawaccording to the following calculation:

${{Alkaloid}\mspace{14mu}{content}\mspace{14mu}(\%)} = {\frac{0.1 \times \frac{{SPLA} \times {STDC}}{STDA} \times ( {{EV} + \frac{ {{LOD}\mspace{14mu}\% \times {SW}} )}{100}} }{{SW} \times \frac{( {100 - {{LOD}\mspace{14mu}\%}} )}{100}} \times \frac{STDI}{SPLI}}$

-   -   where SPLA is the area under the sample peak of interest    -   STDC is the concentration of the standard alkaloid in mg/mL    -   STDA is the area under the standard peak    -   EV is extractant volume in mL    -   LOD % is the loss on drying of the straw, expressed as a        percentage    -   SW is straw weight extracted in grams.    -   STDI is the volume of standard injected in microliters    -   SPLI is the volume of sample injected in microliters

FN1-900-1 had noticeably low vigour compared with the other lines. Thevigour of FN1-1242-3 appeared to be normal. The vigour differencesbecame apparent well after establishment, indicating that it was not aseed quality effect.

Table 6 shows the loss on drying of straw and the mean alkaloid contentof the duplicate straw samples as determined by UPLC.

TABLE 6 Amount of capsule, straw and seed harvested, loss on drying(LOD) of straw, and alkaloid content determined by UPLC. Yield (kg/line)air dry basis Alkaloid content (dry weight basis) Straw/ Straw ThebaineOripavine Thebaine Selection Capsule Seed Straw Capsule ratio LOD (%) %(DWB) % (DWB) Total % ratio FN1-900-1 2.91 1.27 1.64 0.56 7.3 3.07 0.013.08 1.00 FN1-1242-3 3.33 1.22 2.11 0.63 7.8 3.23 0.02 3.25 0.99Thebaine ratio is the thebaine content divided by the sum of thebainecontent and oripavine content.

The results show that the poppy straw in the two lines FN1-900-1 andFN1-1242-3 are very high in thebaine content, and very low (0.01% and0.02%, respectively, in oripavine content). There were no otheralkaloids (i.e., morphine, codeine, salutaridine, reticuline, laudanine,papaverine and noscapine) detected using the method described.

Growing and Evaluation of M4 Generation

Using seed harvested from the M3 generation, 2 large field plots ofFN1-1242-3 and one of FN1-900-1 were grown using commercial equipmentand methods. No growth regulator sprays were used. Table 7 summarisesthe alkaloid contents achieved in these crops. At the first site(Elphinstone, Circular Head district of Tasmania), both selections weregrown, along with the Norman parent line, WF03-0802. Hand-picked samplestaken from the plots assayed at over 5.0% thebaine, and 0.02% oripavine.Machine harvested samples assayed at 4.65% thebaine and 0.02% oripavine(FN1 lines combined). The Norman parent line in the same paddock assayedat 3.05% thebaine and 0.89% oripavine.

At Roebuck's (Merseylea district of Tasmania), the FN1-1242-3 cropassayed at 4.36% thebaine and 0.02% oripavine, whilst the Norman parentline WF03-0802 assayed at an average of 2.08% thebaine and 0.75%oripavine. This data shows that the thebaine-only trait has beeninherited into the M4 generation, and that it enables the plants toaccumulate very high contents of thebaine.

The method used for the straw analysis is as follows. Ten gram samplesof ground straw were extracted with 100 mL extraction solution. Theextraction solution comprised 30% ethanol and 10% glacial acetic acid.The samples were shaken for 25 minutes and then filtered through WhatmanNo. 6 filter paper. The solutions were analysed using a Waters AllianceHPLC system fitted with a Alltech Platinum C18 column, 7 mm×53 mm, with3 micron packing.

The mobile phase consisted of 8 mL triethylamine, 125 acetonitrile and950 mL MilliQ water, adjusted to pH 4.1 with phosphoric acid. The flowrate was 3 mL/minute and the column was maintained at 40 C. Thealkaloids were detected using a UV detector at 284 nm.

The loss on drying (LOD) of the straw was determined by heating a sampleat 88° C. for 9 minutes using an infrared (IR) balance (A&D Company LtdModel AD4717, Japan).

Alkaloid concentrations were determined by comparison with standardsolutions, and results calculated on a dry weight basis. The thebaineand oripavine peaks in these samples accounted for over 98%, andmorphine and codeine accounted for less than 1%, of the alkaloid peakarea, indicating that there was substantially no morphine and codeine inthese samples.

TABLE 7 Summary of commercial results for thebaine-only lines (M4generation), compared with parent line. Grower/Load No Description LineThebaine % Oripavine % Total % Elphinstone 1273 Hand FN1-1242-3 5.100.02 5.12 1274 picked FN1-900-1 5.25 0.02 5.27 samples 1302 Parent lineWF03-0802 3.05 0.89 3.94 control (machine harvested) 1312 FN1 linesFN1-1242-3 4.65 0.02 4.67 combined FN1-900-1 (machine harvested) Roebuck1047 Parent line WF03-0802 2.07 0.77 2.84 control 1060 Parent lineWF03-0802 2.09 0.73 2.82 control 1115 FN1 line FN1-1242-3 4.36 0.02 4.38Growing and Evaluation of M5 Generation

Seeds of the M5 generation of FN1-1242-3 were sown in a field trial atGawler, Tasmania on 29 Aug. 2008, along with seeds of the parent lineWF03-0802. Both lines were sown in three blocked replications in plots 5m long by 1.6 m wide. Standard commercial practices were used to growthe trial. No growth regulator sprays were used. The trial was harvestedon 19 Feb. 2009 by hand picking all the capsules within 2 m² quadratswithin each plot. The samples were threshed and the poppy straw weighed.After grinding to <2 mm, the poppy straw was extracted and analysedusing the same method as used for the M3 generation (described above).Table 8 shows the mean alkaloid contents in the straw and the alkaloidyields per hectare. In this example, the thebaine content of the strawof FN1-1242-3 is 96.3% of the total, and oripavine content is 1.47% ofthe total, where the total is the sum of morphine, codeine, thebaine andoripavine content of the poppy straw.

TABLE 8 Summary of trial results for thebaine-only line FN1- 1242-3 (M5generation), compared with parent line. Alkaloid yield Alkaloid content% (dry weight basis) (kg/hectare) Thebaine Line Morphine CodeineThebaine Oripavine Total Thebaine Oripavine ratio WF03-0802 0.13 0.002.54 1.71 4.38 29.0 19.6 0.60 FN1-1242-3 0.09 0.00 3.92 0.06 4.07 58.60.95 0.98 Total is the sum of the contents of morphine, codeine,thebaine and oripavine. Thebaine ratio is the thebaine content dividedby the sum of thebaine content and oripavine content. Alkaloid yield isobtained by multiplying the alkaloid content by the straw yield.

Example 2 Latex Extraction Reagent

Latex Extraction Buffer: 23 g of ammonium dihydrogen phosphate wasdissolved in approximately 750 mL deionised water and 200 mL of ethanoladded, and made up to 1 L with deionised water.

Method

Isocratic Method:

A Pall AcroPrep™ 96 well, 0.2 μm GHP filter plate was placed on a 96well, 350 μL collection plate. Both filter and collection plate werelabeled and 280 μL of buffer pipetted into each well of the filter plateusing a multipipette. Using forceps, a leaf tip approx 5 mm×5 mm wastorn off from the plant to be tested and added to the extractant. Thelatex will bleed into the solution over time.

The sample was allowed to incubate at room temperature for at least 30minutes. The sample was filtered using a vacuum manifold (PallCorporation product No. 5017). The collection plate was covered withAbgene® adhesive PCR sealing foil (Cat #: AB-0626) to preventevaporation. The collection plate can be stored in the refrigerator orfreezer pending analysis.

Analysis Method

Instrument:

-   -   Waters Acquity UPLC®, with Sample Organiser and Tunable Ultra        Violet (TUV) detector    -   Waters Column, Bridged Ethyl Hybrid (BEH) particles, C18, 1.7        μm, 2.1×50 mm    -   TUV detector, wavelength 284 nm

Reagents:

Mobile Phase A—9% methanol, 0.1% formic acid, adjusted with ammonia topH=9.6

Mobile phase B—91% methanol, 0.1% formic acid, adjusted with ammonia topH=9.6

Weak Wash—10% methanol

Strong Wash—100% methanol

The Sample Manager option “Load Ahead” was used to save time betweensamples. With this option, each sample was aspirated ready for injectionwhile the previous one was running.

TABLE 9 Mobile phase settings Time Flow Rate (min) (mL/min) % A % B0-0.8 0.7 35.0 65.0

The samples were automatically injected (injection volume 2.0 or 3.5 μL)and chromatographed by the Acquity UPLC® along with standard referencealkaloids. After the sample set has been run by the Acquity UPLC®, thepeaks were identified by comparison with the standards that were run inthe sample set. Typical retention times were as follows:

Retention time Alkaloid (minutes) Morphine 0.24 Oripavine 0.27 Codeine0.31 Papaverine 0.38 Thebaine 0.42 Noscapine 0.68

The separations obtained using this method are shown in FIG. 1. Althoughthe peak shapes and separations are not perfect, they are quite adequatefor a very rapid screening method.

Empower software (Waters Corporation, Milford, Mass.) was used toidentify peaks and calculate peak areas. The data was then exported toan Excel spreadsheet where peak area data was used to determine whichpoppies had unusual alkaloid profiles.

Gradient Method:

For more accurate repeat analysis of samples, a 2.5 minute gradient UPLCmethod was used. It is the same as described above, except that thefollowing gradient conditions were used. FIG. 2 provides a chromatogramof the poppy straw of the M3 generation of FN1-1242-3. The injectionvolume of the sample in FIG. 2 was 2.0 μL. ATA indicates thebaine, onindicates oripavine peak, AMA represents morphine, and ACA representscodeine.

TABLE 10 Instrument method details for 2.5 min gradient method Time FlowRate (min) (mL/min) % A % B Curve 0 0.8 75.0 25.0 1.4 0.8 1.0 99.0 6 2.50.8 75.0 25.0 1

Example 3 Determination of Genetics of Thebaine-Only Trait

Crosses were conducted between FN1 lines with low content of oripavine(<2% of combination of morphine, codeine, oripavine and thebaine) andnormal poppy lines containing morphine. Plants of the first F1generation all contained morphine, indicating that the genes responsiblefor the thebaine-only characteristic are recessive. The F1 plants wereself pollinated. Seeds were collected from the F1 plants, and sown intrays. When the plants were at the 6-leaf stage, latex testing wasconducted to determine the chemotypes of the individual F2 plants.

Latex testing was done according to the method of Example 4 by removingthe tip of the youngest fully expanded leaf and placing it in acidextractant buffer in a Pall filter plate. After allowing time for thelatex to bleed from the leaf into the buffer, the extractant was vacuumfiltered into a 96 well plate and sealed. The samples were analysed byUPLC using the method shown in Example 4.

The alkaloid concentrations were calculated from the peak areas byreference to standard alkaloid solutions, and each alkaloid wasconverted to a percentage of total morphinan alkaloid (morphine,codeine, thebaine and oripavine) in order to determine the chemotype ofthe plant. A set of rules incorporated in nested “IF” statements wasestablished to determine chemotype. The rules, applied sequentially,were as follows:

If total concentration of morphine, codeine, thebaine and oripavine isless than 5 ug/mL in injected solution, no result.

If Noscapine percentage >15, chemotype=Noscapine.

If Thebaine percentage >98, chemotype=Thebaine-only.

If Thebaine+Oripavine percentage >95, chemotype=Norman.

If Thebaine+Codeine percentage >96, chemotype=Codeine.

If Morphine percentage >2, chemotype=Morphine.

Otherwise, the chemotype was classified as OCT, which indicates that theplant contained oripavine, codeine and thebaine.

Four chemotypes were identified in the populations:

Morphine: morphine present typically with thebaine and codeine

Norman: thebaine and oripavine both present, substantially no morphine

Thebaine-only: thebaine present, substantially no oripavine and morphine

Codeine: thebaine and codeine present, substantially no morphine

The occasional plant was identified as OCT. These generally were verysmall plants that fitted into one of the four categories as they furtherdeveloped. Overall, 0.18% of plants were classified as OCT and 0.03%were classified as noscapine and were ignored in calculation of ratiosfor this analysis.

Chi square tests were conducted to determine if the observed segregationpatterns differed significantly from a 9:3:3:1 segregation of chemotypes(Morphine: codeine: Norman: Thebaine-only respectively).

Table 11 shows the results of the 9 populations derived from FN1 parentshaving a thebaine-only chemotype. Five of these fit a 9:3:3:1 ratio(P<0.05). Of those that didn't fit the expected ratio, FN1-1242 isnotable, in that it had more than the expected number of plants with thecodeine chemotype, whilst all other lines that did not fit the ratio hadless than expected codeine plants. Further work was conducted withprogeny of FN1-1242 in which plants were grown to hook stage, and leaflatex was collected and classified into chemotypes. The distribution was96:41:42:11, which gives a Chi square test of 3.1 which indicates thatthere is no significant difference from the expected ratio.

The segregation into 4 chemotypes indicates that two separate genes areinvolved in the thebaine-only chemotype. One of these is the geneassociated with the Norman poppy mutation (described in U.S. Pat. No.6,067,749). The second gene is responsible for the new low-oripavine orthebaine-only trait, which is responsible for blocking the pathwaybetween thebaine and oripavine. These two genetic changes work togetherin lines described herein to provide the poppy plants with a highthebaine content and a low oripavine content.

TABLE 11 Segregation patterns for populations using FN1 lines with <2%oripavine in alkaloid profile. Alkaloid content in straw of FN1 Observedratio Chi square test result M2 plant as percentage of sum ofMorphine:co- (ns indicates no morphine, codeine, oripavine anddeine:Nor- significant thebaine man:thebaine- difference from FN1 lineOripavine % Thebaine % only 9:3:3:1 ratio) FN1-846-2 0.24 98.439:14:11:4 0.39 ns FN1-1719-1 0.31 99.5 45:10:1:0 17.9 < 0.001 FN1-21990.38 99.6 59:25:18:6  1.5 ns FN1-1242 0.41 99.2 271:112:81:16 12.81 <0.01  FN1-900 0.58 98.8 191:73:55:19 2.79 ns FN1-2152 1.38 98.673:2:28:14 26.7 < 0.001 FN1-1571 1.63 97.9 604:58:248:69 114.7 < 0.001 

Example 4 Leaf Latex Analysis Reagent

Acid Extractant: A 1 L measuring cylinder was half filled with deionisedwater. 1 mL of conc. phosphoric acid and 50 mL ethanol were added andthe volume made up to 1 L with deionised water.

Method

A Pall AcroPrep™ 96 well, 0.2 μm GHP filter plate was placed on a 96well, 350 μL collection plate. Both filter and collection plate werelabeled and 280 μL of acid extractant pipetted into each well of thefilter plate using a multipipette. A tip of the youngest fully expandedleaf was torn off each plant to be tested and added to the extractant.The latex bleeds into the solution over time.

The samples were allowed to incubate at room temperature for at least 30minutes. The samples were filtered using a vacuum manifold (PallCorporation product No. 5017). The collection plate was covered withABgene® adhesive PCR sealing foil (Cat #: AB-0626) to preventevaporation. The collection plate can be stored in the refrigerator orfreezer pending analysis.

Analysis Method

The samples were analyzed using the same instrument, reagents andinstrument method details (see Table 10) as described for the gradientmethod in Example 2.

The samples were automatically injected and chromatographed by theAcquity UPLC® along with standard reference alkaloids. After the sampleset has been run by the Acquity UPLC®, the peaks were identified bycomparison with the standards that were run in the sample set. Empowersoftware (Waters Corporation, Milford, Mass.) was used to identify peaksand calculate peak areas. The data was then exported to an Excelspreadsheet where peak area data was used to determine alkaloidprofiles.

While the foregoing specification teaches the principles of the presentinvention, with examples provided for the purpose of illustration, itwill be understood that the practice of the invention encompasses all ofthe usual variations, adaptations and/or modifications as come withinthe scope of the following claims and their equivalents.

What is claimed is:
 1. A poppy straw comprising a poppy straw of astably reproducing Papaver somniferum having thebaine constituting about95% by weight or greater of an alkaloid combination, and havingoripavine constituting about 5% by weight or less of the alkaloidcombination, wherein the alkaloid combination comprises morphine,codeine, thebaine and oripavine; and wherein thebaine constitutes about3.0% or greater of the poppy straw on a dry weight basis.
 2. The poppystraw of claim 1, wherein thebaine constitutes about 3.5% or greater ofthe poppy straw on a dry weight basis.
 3. The poppy straw of claim 1,wherein thebaine constitutes about 4.0% or greater of the poppy straw ona dry weight basis.
 4. The poppy straw of claim 1, wherein thebaineconstitutes about 97% by weight or greater of the alkaloid combinationand oripavine constitutes about 0.7% by weight or less of the alkaloidcombination.
 5. The poppy straw of claim 1 wherein there issubstantially no morphine or codeine in the alkaloid combination.
 6. Thepoppy straw of claim 5, wherein there is substantially no oripavine inthe alkaloid combination.
 7. A plant comprising a stably reproducingplant of Papaver somniferum which upon the harvesting of its poppycapsules will yield a poppy straw having thebaine constituting at least3% by weight on a dry basis, and oripavine constituting no more than0.4% by weight on a dry basis of said straw.
 8. The plant of claim 7having thebaine constituting at least 3.5% by weight on a dry basis ofsaid straw.
 9. The plant of claim 7 having thebaine constituting atleast 4.0% by weight on a dry basis, and oripavine constituting no morethan 0.2% by weight on a dry basis of said straw.
 10. A seed of theplant of claim
 7. 11. Papaver somniferum seeds which have been depositedunder ATCC accession number PTA-9109.
 12. A Papaver somniferum planthaving a high thebaine content of over 3% in the straw on a dry weightbasis wherein the high thebaine content is provided by two independenttraits, one trait controlling the accumulation of thebaine and oripavinecompared with morphine and codeine, and the second trait controlling theaccumulation of thebaine compared with oripavine.
 13. A Papaversomniferum plant having a high thebaine content of over 3% in the strawon a dry weight basis wherein the high thebaine content is provided bytwo independent genetic changes, one genetic change controlling theaccumulation of thebaine and oripavine compared with morphine andcodeine, and the second genetic change controlling the accumulation ofthebaine compared with oripavine.
 14. An opium for the extraction ofthebaine comprising an opium of the plant of claim
 7. 15. A concentrateof poppy straw for the extraction of thebaine comprising a concentrateof poppy straw of the plant of claim
 7. 16. A method for the productionof thebaine which comprises the steps of: a) harvesting poppy capsulesof the plant of claim 7 to produce a poppy straw; and b) chemicallyextracting the thebaine from the poppy straw.
 17. A method for theproduction of thebaine which comprises the steps of: a) collecting anddrying the latex of the immature poppy capsules of the plant of claim 7to produce opium; and b) chemically extracting the thebaine from theopium.
 18. A stand of stably reproducing Papaver somniferum plants ofclaim
 7. 19. The poppy straw of claim 1, wherein oripavine constitutesabout 0.4% or lower of the poppy straw on a dry weight basis.
 20. Thepoppy straw of claim 1, wherein oripavine constitutes about 0.2% orlower of the poppy straw on a dry weight basis.
 21. The poppy straw ofclaim 20, wherein oripavine constitutes between 0.01 to 1.0% by weightof the alkaloid combination.
 22. The poppy straw of claim 21, whereinoripavine constitutes between 0.02% to 0.5% by weight of the alkaloidcombination.
 23. A poppy straw comprising a poppy straw of the plant ofclaim
 7. 24. A method for producing a poppy plant of Papaver somniferumhaving a stably heritable high thebaine content and low oripavinecontent versus morphine and codeine content, the method comprising thesteps of: a) exposing at least one poppy seed of Papaver somniferum to amutagenizing agent, b) growing the at least one poppy seed to produce aplant bearing a leaf or an immature poppy capsule, optionally throughmultiple self-fertilized generations, c) sampling the leaf or poppycapsule for presence of thebaine, oripavine, morphine and codeine, andd) repeating steps b) and c) and optionally step (a) until a poppy plantof Papaver somniferum is obtained which yields a poppy straw havingthebaine constituting about 95% by weight or greater of the alkaloidcombination consisting of morphine, codeine, thebaine and oripavine, andhaving oripavine constituting about 5% by weight or less of the alkaloidcombination, and wherein thebaine constitutes about 3.0% or greater ofthe alkaloid content of the poppy straw on a dry weight basis.
 25. Themethod according to claim 24 wherein the Papaver somniferum seed exposedto the mutagenizing agent in step (a) is a Papaver somniferum yielding,upon the harvesting of its poppy capsules, a poppy straw having athebaine and oripavine content constituting 50% by weight or greater ofthe alkaloid combination consisting of morphine, codeine, thebaine andoripavine.
 26. The method according to claim 24 wherein step (b)comprises growing the at least one poppy seed to produce a plant bearinga leaf or an immature poppy capsule, and self-pollinating to produceseed, and taking the seed thereby produced and producing an M2generation of plants, and step (c) comprises screening the M2 plants andselecting plants which yield a poppy straw having thebaine constitutingabout 95% by weight or greater of the alkaloid combination consisting ofmorphine, codeine, thebaine and oripavine, and having oripavineconstituting about 5% by weight or less of the alkaloid combination, andwherein thebaine constitutes about 3.0% or greater of the poppy straw ona dry weight basis.
 27. The method according to claim 25 wherein thePapaver somniferum seed exposed to the mutagenizing agent in step (a) isseed selected from the group consisting of ATCC PTA-9110 and ATCCPTA-9109.
 28. Progeny of Papaver somniferum ATCC-9109, said progenyyielding a poppy straw having thebaine constituting about 95% by weightor greater of an alkaloid combination, and having oripavine constitutingabout 5% by weight or less of the alkaloid combination, and whereinthebaine constitutes about 3.0% or greater of the alkaloid content ofthe poppy straw on a dry weight basis.
 29. A mutant or variant of aPapaver somniferum selected from the group consisting of ATCC PTA-9109and ATCC PTA-9110, said mutant or variant yielding a poppy straw havingthebaine constituting about 95% by weight or greater of an alkaloidcombination, and having oripavine constituting about 5% by weight orless of the alkaloid combination, and wherein thebaine constitutes about3.0% or greater of the alkaloid content of the poppy straw on a dryweight basis.